Fig. 6

VMP1 palmitoylation affects SSCLC self-maintenance and proliferation in co-culture. A Schematic diagram of co-culture of SSCLCs with SCs transfected by WT-VMP1-Flag, MT-VMP1-Flag or NC plasmid, with SSCLCs in the lower chamber and SCs in the upper chamber of a Transwell (0.4 μm). B SCs-derived sEVs were labeled by PKH26 dye and then incubated with SSCLCs for 24 h. Representative fluorescence images of PKH26-labelled (red) SSCLCs. The control group was treated with the medium of dye solvent without PKH26. C SCs were transfected with WT-VMP1-Flag, MT-VMP1-Flag or NC plasmid respectively in the upper layer. After 24 h of co-incubation, the proportion of PLZF positive cells in the lower layer was assessed by flow cytometry to represent SSCLC ratio in WT-VMP1, MT-VMP1, NC groups on the 12th day of induction. D Real time qPCR experiment showed the expression of SSC-related genes PLZF, GFRα1, ID4 and spermatogonial related genes DMRT1 in WT-VMP1, MT-VMP1 and NC groups. E The proliferation rate of SSCLCs was tested by EDU assay in WT-VMP1, MT-VMP1 and NC groups. F The apoptosis rate of SSCLCs was detected by FITC-Annexin V flow cytometry in WT-VMP1, MT-VMP1 and NC groups. Black boxes represent the apoptosis ratio of SSCLCs. Three independent times of each experiment were performed and statistically analyzed. Data were presented as mean ± SEM, *P < 0.05