TSP50 promotes hepatocyte proliferation and lipid metabolism by mediating G6PD activity. A‐C, pcDNA3.1‐TSP50 in combination with Flag‐G6PD, Flag‐G6PD K171Q or Flag‐G6PD K403Q was co‐transfected into L02, Huh7 and Bel7402 cells to overexpress TSP50 and G6PD wild‐type or mutants’ vectors. The efficient expression of TSP50 or G6PD (wild type and mutants) was examined by Western blot analysis using anti‐TSP50 and anti‐Flag antibodies, respectively. D‐E, TG and T‐CHO contents were detected in cells that co‐transfected with TSP50 and G6PD or G6PD K171Q or G6PD K403Q. F‐H, Relative expression of lipid metabolism–related genes was detected by RT‐PCR assay. I‐K, Cell viability was detected by MTT assay after co‐transfection of pcDNA3.1‐TSP50 with Flag‐G6PD or Flag‐G6PD mutants in L02, Huh7 and Bel7402 cells. L‐N, The BrdU incorporation assay was performed in cells after co‐transfection of pcDNA3.1‐TSP50 with Flag‐G6PD or Flag‐G6PD mutants. Data were presented as mean ± SD. *P < .05 as compared to NC group by one‐way ANOVA. **P < .01