Research Paper Volume 11, Issue 19 pp 8156—8168

Knockdown of CREB3 activates endoplasmic reticulum stress and induces apoptosis in glioblastoma

Figure 4. The downregulation of CREB3 induced apoptosis in U251MG cells. (A) CREB3 levels were detected by qRT-PCR in U251MG cells transfected with the NC, CREB3-shRNA1 or CREB3-shRNA1. (B, C) CREB3 levels were measured by Western blotting in U251MG cells transfected with the NC, CREB3-shRNA1 or CREB3-shRNA2. β-actin was used as a loading control. (D, E) U251MG cells were transfected with CREB3-shRNA2 for 72 h. Apoptosis was detected with Annexin V and PI double staining, and the percentage of apoptotic cells was calculated. (F) BAX and active caspase 3 levels were detected by Western blotting in U251MG cells transfected with CREB3-shRNA2 for 72 h. (G) The relative levels of BAX and active caspase 3 were quantified and normalized to β-actin levels. *P<0.05, **P<0.01 compared with the NC group.