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. 2013 Jul 20;7(6):1012–1018. doi: 10.1016/j.molonc.2013.07.010

Figure 2.

Figure 2

Transcriptional regulation of SESN2 by ER stress and ATF4. A) HeLa cervical cancer cells were treated with ER stress‐inducing drug thapsigargin (THA, 100 nM) for the indicated periods and analyzed by RT‐PCR for the expression of SESN2 and the expression and splicing of ER stress‐regulated transcription factors. B) HeLa cells were transfected with expression plasmids for ATF4 (pcDNA3.1‐ATF4‐V5), ATF3 (pcDNA3.1‐ATF3‐V5), and CHOP (pcDNA3.1‐CHOP‐V5), and analyzed for the expression of SESN2 by RT‐PCR analysis. Since ATF4 protein expression is mainly regulated at the posttranscriptional level and transfected ATF4 mRNA cannot be distinguished from endogenous mRNA, a combination PCR of CMV promoter‐specific primers and ATF4 reverse primers was used to prove for high transfection efficiency.