Fig. 3.

CRISPR-Cas9 KO screening identifies POLRMT as a potential target to enhance the efficacy of DAC. Thirteen mtRNA regulators were screened using the CRISPR-Cas9 system. Individual polyclonal KO cells were treated by DMSO or DAC and the cell proliferation was analyzed by CCK-8 assay. (B) RT-qPCR analysis of mt-mRNAs in POLRMT KO cells. (C) Protein expression of POLRMT and COX1 in POLRMT KO cells was analyzed by western blotting. (D) Glycolytic and mitochondrial ATP content in control and POLRMT KO cells treated with DMSO or DAC. (E, F) RT-qPCR analysis of TCF21 (E) and several ISG (F) mRNA levels in control and POLRMT KO cells treated with DMSO or DAC. In all RT-qPCR experiments, the quantification of transcript level is relative to a control (ACTB). In all experiments, an average of three biological replicates is shown with error bars denoting s.e.m. A Student’s t-test was performed for statistical analysis. * indicates P-value < 0.05, ** indicates P-value < 0.01, *** indicates P-value < 0.001.