Venice Aztec Print Sleeveless Top In Yellow The Fashion Bible Buy Cheap Exclusive Cheap View FPKcwqR0

Venice Aztec Print Sleeveless Top In Yellow The Fashion Bible Buy Cheap Exclusive Cheap View FPKcwqR0
Venice Aztec Print Sleeveless Top In Yellow The Fashion Bible
Agency Links Links
Main Menu
All Categories
Gov. Eric J. Holcomb
Eric J. Holcomb
Visit His Homepage »

To construct the hotspot luciferase reporters, a Myogenin minimal promoter ( 16 ) was amplified from C2C12 genomic DNA and inserted into pGL3 basic vector (Promega) using Xho I and Hind III sites. Individual hotspot was then cloned into the downstream of the firefly luciferase gene using BamH I and Sal I sites. For generating the reporters with hotspot combination and deleting MyoD binding motif on H3 or H4 reporter, fusion polymerase chain reaction (PCR) was used by employing overlapping annealing sequences as described before ( 17 ). HA-tagged MyoD expression plasmids were kind gift from Dr Slimane Ait-Si-Ali. Flag-tagged FoxO3 expression plasmid was a kind gift from Dr Ping Hu.

All the cells were transfected using Lipofectamine 2000 (Life Technologies). The Renilla plasmid (Promega) was co-transfected as a normalization control. At 24 h post-transfection, C2C12 cells were differentiated for 48 h and the luciferase activity was measured using the Dual-Glo Luciferase Assay system (Promega) according to the manufacturer's guidelines.

ChIP assays were performed as previously described ( 18 , 19 ). For ChIP, C2C12 cells were crosslinked with 1% formaldehyde at room temperature for 10 min. For sequential ChIP, cells were double crosslinked with 1 mM DSG for 45 min and then for 10 min by 1% formaldehyde. In both cases, crosslinking reaction was quenched by addition of 0.125M glycine for 10 min. Chromatin was fragmented using sonicator, followed by incubation with 5 μg of antibodies at 4°C for overnight. Antibodies for MyoD (Santa Cruz Biotechnology, sc-304X, rabbit polyclonal), FoxO3 (Santa Cruz Biotechnology, sc-48348X, rabbit polyclonal) and histone H3-K27 acetylation (Abcam, ab4729, rabbit polyclonal) were used for ChIP, or normal rabbit IgG (Santa Cruz Biotechnology, sc-2027) was used as negative control. For sequential ChIP, the first ChIP was performed as described above and the immunocomplexes were eluted with 50 μl of 10 mM Dithiothreitol (DTT) at 37°C for 30 min. The eluted supernatant was diluted for 20 times, followed by incubation with antibodies for the second ChIP or IgG. Immunoprecipitated genomic DNA was resuspended in 50 μl of water. PCRs were performed with 1 μl of immunoprecipitated DNA as template with SYBR Green Master Mix (Life Technologies) and products were analyzed by qRT-PCR on a 7900HT system (Life Technologies). Primers used are listed in Long Sleeve ButtonDown Shirt With Eyelet Embroidery Blue Derek Lam Exclusive For Sale Discount Sale Aaa Quality Sale Order Store Cheap Online qXsLyirPt

Chromosome conformation capture (3C) was performed as previously described ( 20 ). Briefly, 2 × 10 7 C2C12 cells were fixed by adding 2% formaldehyde at room temperature for 10 min, followed by quenching the crosslinking reaction by adding 0.125 M glycine for 10 min. Cells were lysed with lysis buffer (10 mM Tris–HCl pH 8.0, 10 mM NaCl, 0.2% Igepal CA630, with protease inhibitor cocktail freshly added) rocking on ice for 10 min. Next, nuclei were centrifuged and re-suspended in CutSmart Buffer (NEB). The extracted chromatin was then digested with 400 units of EcoRI (NEB). On the next day, digested chromatin was ligated by 100 units of T4 DNA Ligase in 10 times of the initial volume at 16°C for 4 h. The chromatin was subsequently de-crosslinked at 65°C for overnight. On the next day, protein was removed by incubating with 300 μg of protease K (Life Technologies) at 55°C for 2 h, and RNA was removed by 300 μg of RNaseA (Life Technologies) at 37°C for 2 h. DNA was extracted by phenol/chloroform isolation, finally resuspended in 250 μl of 1× TE buffer. The digestion and ligation efficiencies were assessed before performing PCR. Probes were designed near enhancers and promoter. PCRs were performed with BAC clone RP24–343C13 as control, with SYBR Green Master Mix (Life Technologies) and products were analyzed by qRT-PCR on a 7900HT system (Life Technologies). The sequence and layout of probes were listed in Supplementary Data S1 .

DESIGN high neck jumper in fine knit Lilac Asos Sale Best Seller GQdKi9g
| 6 min read

Written by Erik Devaney

Marketing | 6 min read
Share to Twitter Share to Facebook Share to Email App Share to LinkedIn Share to Messenger Share to Slack

The Dark Ages.

For those of you unfamiliar with Medieval history, the Dark Ages refer to a period of crippling technologicaldegradation. A time when there was no No Netflix. No Hulu. No Spotify or Pandora or Sirius radio.

During the Dark Ages,consumersdidn’t get personalized recommendations based on their past purchases. Nor did they receive personalized recommendations for new music, movies, or TV shows based on what they’d already listened to and watched.

In fact, to watch their favorite TV shows, people had to“tune in” at specific times. Presumably, the TV industry was creating content for us . Yet if we ever wanted to watch it, we had to do so on their schedules.

It was truly a dark time.

OK, so my Medieval history might be a little off. But my underlying point here is that people prefer -- and often crave -- personalized experiences. And by“personalized experience,” I mean an interaction or engagement with apiece of software, a piece of content, or a person (duh) that leaves you feeling like your interests and preferences were actually being taken into account.

Personalization is like someone giving you a fitted baseball cap with your favoriteteam’s logo on the front and yourinitials stitched in on the side.

In contrast, non-personalization is like someone giving you a one-size-fits-all baseball cap with some team you hate’s logo on the front. No initials. No consideration for your preferences whatsoever. It's like the person who gave it to you bought a 48-pack ofbaseball caps on Amazon and you were just one of the many “lucky" recipients.

If you’d like to learn more about incorporating personalization into your marketing, check our new guide, Online Cheap Cheap Enjoy Romy ballerinas Nude amp; Neutrals Jimmy Choo London Original puWbKXmpL

How to Master Personalized Marketing

If, however, you want to learn more about why people crave personalization at a psychological level, just keep on reading and I’ll do my best to explain.

Why Do We Prefer Personalized Experiences?

According to a study from the University of Texas , we canattribute our preference for personalized experiences to two key factors: desire for control and information overload.Let’s tackle“desire for control" first.

So, we know that a personalized experience -- by its very nature -- is in some way different from the status quo. You’re not just getting what everyone else is getting with personalization. Instead, you’re getting something tailored to you. And because of that, it makes you feel more in control.

Advanced Search
Article navigation
Volume 28
Issue 3
December 2001
Darren W. Dahl
Search for other works by this author on:
Oxford Academic
Google Scholar
Rajesh V. Manchanda
Search for other works by this author on:
Oxford Academic
Google Scholar
Jennifer J. Argo
Search for other works by this author on:
Oxford Academic
Google Scholar
, Volume 28, Issue 3, 1 December 2001, Pages 473–481,
01 December 2001
Article history
01 September 1999
Revision Received:
01 March 2001

Darren W. Dahl, Rajesh V. Manchanda, Jennifer J. Argo; Embarrassment in Consumer Purchase: The Roles of Social Presence and Purchase Familiarity, Journal of Consumer Research , Volume 28, Issue 3, 1 December 2001, Pages 473–481,

Download citation file:

© 2018 Oxford University Press

Advanced Search


Two field studies investigate the importance of social presence (real and imagined) and familiarity with the purchase act in producing embarrassment in the context of an embarrassing product purchase. The results indicate that awareness of a social presence during purchase selection and commitment, whether real or imagined, is a motivating factor in creating embarrassment for the consumer. Further, our results show that the more familiar consumers are with an embarrassing product purchase, the less embarrassed they are likely to feel. Familiarity with an embarrassing product purchase is also shown to have implications for the effect of social presence. That is, familiarity with purchase acts as a moderator for the relationship of real social presence and embarrassment by reducing the influence of the social presence. In the context of an imagined social presence, purchase familiarity is shown to reduce the likelihood of imagining. These findings are integrated into a discussion of the theoretical implications and the potential avenues for future research in the area.

Previous studies of the etiological role of psychological stress and the occurrence of coronary artery disease have almost exclusively focused on chronic risk and have yielded controversial results. 7 Recent studies suggest that hostility, cynicism, and anger comprise a critical “toxic” component 8 of type A behavior and are more strongly associated with the incidence of coronary artery disease than other aspects of global type A behavior. 7 8 9 Jane mandarincollar cottondenim shirtdress APC Cheap Fashionable Gy4vG4I1
We therefore chose to focus on discrete episodes of anger as a potential trigger of acute myocardial infarction. Clarification of the role of anger as a trigger is important because a better understanding of triggering could facilitate approaches to sever the link between potential triggering activities and the transient physiological risk states that produce myocardial infarction.

The Determinants of Myocardial Infarction Onset Study (Onset Study) is a multicenter, interview-based study in patients with acute myocardial infarction. Discount With Mastercard Free Shipping Best Seller Boatneck Boyfriend Tee underwater pink heart by VIDA VIDA Wiki Online lGMdFGRJA
In this study, we used a case-crossover design 12 13 to quantify the relative risk of myocardial infarction onset after discrete episodes of anger in 1623 patients with confirmed acute myocardial infarction.

Study Population

The Onset Study was conducted in 22 community hospitals and 23 tertiary care centers (see “Appendix”). Between August 1989 and March 1993, 1623 patients (1122 men and 501 women; age range, 20 to 92 years) were interviewed a median of 4 days (range, 0 to 30) after myocardial infarction.

Interviewers identified eligible cases by reviewing coronary care unit admission logs and patients’ charts. For inclusion, patients were required to meet all of the following criteria: (1) elevated creatine kinase level, with positive MB isoenzymes, (2) an identifiable onset of pain or other symptoms typical of the onset of infarction, and (3) ability to complete a structured interview. The protocol was approved by the institutional review board at each participating center, and informed consent was obtained from each patient.

Interviewers were trained by personal instruction, a training manual, an instructional videocassette, and through ongoing feedback from the study coordinator. Approximately one third of the interviews were audiotaped for randomly selected quality control checks of the coding accuracy. To minimize bias in information ascertainment, the interviewers were not informed of the duration of the hypothesized hazard period.

The interview identified the time, place, and quality of myocardial infarction pain and other symptoms, the estimated usual frequency of exposure during the previous year, and the intensity and timing of exposure during the 26 hours before the onset of pain for episodes of anger and other potential triggers of the onset of myocardial infarction. Details pertaining to exposure to heavy physical exertion have been reported elsewhere. 11

Big Sale Cheap Price Womens ContrastNeck Floral Silk Jacquard Blouse Maison Mayle Shop For Sale Clearance Original Cheap Pay With Visa The Cheapest Sale Online TBeAnAhbv

ICF, 530 Gaither Road, Suite 500, Rockville, MD 20850 Tel: +1 301 407-6500 * Fax: +1 301 407-6501

The information provided on this Web site is not official U.S. Government information and does not represent the views or positions of the U.S. Agency for International Development or the U.S. Government.