Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2016 Oct 24;215(2):231–243. doi: 10.1083/jcb.201602001

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2016 Monaco et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — Nuclear calcium signaling differentially regulates CREB activation and CREM expression in primary human T cells. (A, left) Immunoblot analysis of CREB and the serine 133–phosphorylated form of CREB (pCREB) in T cells transfected with expression vectors for CaMBP4.GFP (CaMBP4) or GFP and stimulated with a combination of PMA and ionomycin for 30 or 120 min. A representative example is shown. (right) Quantitative analysis of three experiments; fold induction is relative to the pCREB/CREB ratio in unstimulated GFP-expressing T cells. (B) QRT-PCR analysis of Crem mRNA in T cells transfected with expression vectors for CaMBP4.GFP (CaMBP4) or GFP. Cells were stimulated with a combination of PMA and ionomycin for 6 h or were left unstimulated (ctr). mRNA levels are expressed relative to the mRNA level in GFP-expressing T cells stimulated with a combination of PMA and ionomycin, which was set to 100%. n = 3. Statistically significant differences are indicated with an asterisk (*, P < 0.05). (C and D) FACS analysis of the percentage of IL-2– and CD25-expressing T cells transfected with GFP-tagged expression vectors for either wild-type CREB (wt-CREB) or mutant CREB containing a serine-to-alanine mutation at aa 133 (mut-CREB). Cells were stimulated with a combination of PMA and ionomycin for 6 h or were left unstimulated (ctr). n = 3. Statistically significant differences are indicated with asterisks (*, P < 0.05). (E and F) FACS analysis of the percentage of IL-2– and CD25-expressing T cells. Cells were stimulated with a combination of PMA and ionomycin for 6 h or were left unstimulated (ctr). n = 3. Pretreatment with KN93 was done as indicated. Statistically significant differences are indicated with asterisks (*, P < 0.05). Error bars represent SEM.