Figure 6.

YWHAZ downregulation sensitizes cells to chemo/radiotherapy and suppresses cell growth. (A) Western blotting was performed to detect changes of YWHAZ levels in T24 cells treated with different shRNAs using β‐actin levels as the internal controls. RT‐qPCR was performed to detect mRNA levels of (B) YWHAZ in T24 cells treated with shRNAs and (C) the related apoptosis genes in shRNA‐1‐treated T24 cells. Cells treated with ‘scramble’ served as the control in panels A to C. (D) T24 cells transfected with shRNA‐1 were treated with the indicated concentrations of doxorubicin or cisplatin. Cell viability was measured by Alamar Blue assays 4 days after drug treatments. (E) T24 cells transfected with shRNA‐1 were treated with 6 Gy radiation, after which cell viability was assessed every 24 h for 4 days. (F) Incidence rate of preapoptosis among treated cells 48 h after chemo−/radio‐therapy was determined by annexin‐V staining. (G) Proliferation of shRNA‐1‐treated cells was monitored for 4 days under the treatment of 50 nm doxorubicin or 5 μm cisplatin. (H) Colony formation activity was studied on shRNA‐1‐treated T24 cells after 2 weeks treatment with 50 nm doxorubicin or 5 μm cisplatin (left). Colony numbers per 35‐mm dish are shown in bar charts (right). Cells treated with ‘scramble’ served as the controls in panels D to H. Data were expressed as means ± SD from five replicates in each experimental group. Paired t‐tests were performed to evaluate differences between ‘scramble’‐ and shRNA‐1‐treated cells. *p < 0.05, **p < 0.01, ***p < 0.001.