Figure 4

Suppression of miR-199a-5p promoted sustained ER stress and apoptosis in DCA and TG treated hepatocytes. (a and b) Quantitative PCR analysis of GRP78 and CHOP mRNA levels in miR-199a-5p inh- or scrambled RNA (NC)-transfected hepatocytes after TG, DCA, or DMSO treatment for the indicated times. The mRNA levels were normalized to GAPDH in each sample. Data are shown as means±S.D. of three independent experiments. **P<0.01. (c) Western blotting analysis of GRP78 and CHOP proteins in miR-199a-5p inh- or scrambled RNA (NC)-transfected hepatocytes HL-7702/L02 (short to L02) after DCA or TG treatment for the indicated times. Data are shown as the level normalized to GAPDH in each sample. (d) Apoptotic rates were assayed by flow cytometry in miR-199a-5p inh- or scrambled RNA (NC)-transfected hepatocytes after DCA or TG treatment for 48 h. Data are representative of three independent experiments. (e) Cell death levels of siRNAs-(si-) or NC-RNA-transfected HL-7702/L02 cells were assayed by LDH release after DCA or TG treatment for 48 h. Data are shown as means±S.D. of three independent experiments. **P<0.01, ref to NC-RNA-transfected cells. (f) Cell death levels of miR-199a-5p inh- or NC-RNA-transfected HL-7702/L02 cells were assayed by LDH release after DCA or TG treatment for 48 h. The efforts of miR-199a-5p inh were also rescued by siRNAs targeting to GRP78, ATF6 or IRE1α with scramble RNA as a control. Data are shown as means±S.D. of three independent experiments. **P<0.01