Fig. 6

Knockdown of HDAC4 radiosensitizes EC KYSE30 cells via upregulation of miR-146a and downregulation of IRAK1. Radioresistant KYSE30 (KYSE30R) cells were treated with shRNA (sh-) against HDAC4 and exogenous miR-146a inhibitor alone or in combination under 8 Gy X-ray irradiation. A HDAC4 and IRAK1 expression in radioresistant KYSE30 cells was determined using Western blot analysis, normalized to GAPDH. B miR-146a expression in radioresistant KYSE30 cells was determined using RT-qPCR, normalized to U6. C Colony-formation ability of irradiated (8 Gy)/unirradiated (0 Gy) radioresistant KYSE30 cells was detected by colony formation assay. D Apoptosis of irradiated (8 Gy)/unirradiated (0 Gy) radioresistant KYSE30 cells was detected by flow cytometry based Annexin V-FITC/PI apoptosis assay. E Expression of cleaved-caspase3, cleaved-caspase8, pro caspase3 and pro caspase8 in irradiated (8 Gy)/unirradiated (0 Gy) radioresistant KYSE30 cells was measured using Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate were expressed as mean ± standard deviation. Values among three or more groups by one-way ANOVA followed by Tukey's post hoc test. * p < 0.05 compared with irradiated (8 Gy)/unirradiated (0 Gy) radioresistant KYSE30 cells treated with scramble shRNA. ^#p < 0.05 compared with irradiated (8 Gy)/unirradiated (0 Gy) radioresistant KYSE30 cells treated with sh-HDAC4