Figure 5. Silence of CREB activates expression of αB-crystallin in αTN4-1 cells. (A). CREB knockout strategy in αTN4-1 cells. The homozygous point deletion was created by CRISPR/CAS9 technology. A single nucleotide was deleted in exon 5, and the deletion mutation was verified by DNA sequencing. (B) Western blot analysis of the expression levels of CREB and αB-crystallin in αTN4-1 mock knockdown and CREB knockdown cells. (C, D) Semi-quantification of the western blot results in (B). (E) Apoptosis rate in, mock KO and αTN4-1 CREB KO cells under treatment of 40 mU GO for 6 hours measured by CellTiter-Glo® Luminescent Cell Viability Assay analysis . All experiments were repeated three times. Error bar represents standard deviation, N=3. * p<0.05, *** p<0.005.