Fig. 7: MTCH2 knockout impairs mitochondrial function and inhibits CRPC cell progression.

The single stable pPC-1 cells with a Cas9 construct and the CRISPR/Cas9-MTCH2 KO construct (koMTCH2-cln1, or koMTCH2-cln2, representing two single-cell colonies) or the CRISPR/Cas9-KO vector (“koC”) were cultured and the expression levels of MTCH1 and MTCH2 were assessed using Western blotting (A). The equal number of cells were cultured for designated time periods, the mitochondrial complex I activity (B), ATP contents (C) were tested. Mitochondrial depolarization was determined by JC-1 green monomer intensity (D), and the ROS levels were measured via quantifying CellROX fluorescence intensity (E); Cell proliferation (measured by the percentage of EdU-incorporated nuclei, F), in vitro cell migration (analyzed using “Transwell” assays, G) and apoptosis (measured via nuclear TUNEL staining, H) were tested. The data are expressed as mean ± standard deviation (SD, n = 5). Statistical significance is marked by *P < 0.05 when compared to “koC” cells. The experiments depicted in this figure were conducted five times (biological replicates), consistently producing similar results. The scale bar represents 100 μm.