Fig. 5: Targeting JOSD2 poses a more potent anti-tumor effect on KRAS-mutant CRC over that of KRAS wild-type in vitro/in vivo.
a, b JOSD2 depletion inhibits the cell proliferation of KRAS-mutant cells strongly than KRAS wild-type cells in vitro. HT-29KRAS-WT/Mut cells were infected with indicated lentiviruses for 96 h, then these cells were seeded with 2000 cells per well in 96-well/6-well plates, followed by SRB staining. Ctrl, control. a The representative cells proliferation images of the indicated group; Scale bar, 6 mm. b The quantitative analyses of cells proliferation, P = 0.0965, WT-Ctrl vs. WT-shJOSD2; P = 4E-8, G12C-Ctrl vs. G12C-shJOSD2; P = 6E-8, G12D-Ctrl vs. G12D-shJOSD2; P = 1E-6, G12V-Ctrl vs. G12D-shJOSD2. (means ± SD, n = 3; three independent experiments were performed). c, d JOSD2 depletion inhibits the colony formation of KRAS-mutant cells strongly than KRAS wild-type cells in vitro (means ± SD, n = 3; three independent experiments were performed). c The representative colony formation images of the indicated group; Scale bar, 5 mm. d The quantitative analyses of colony formation. e–h JOSD2 depletion posed a more potent anti-tumor effect on KRAS-mutant xenografts compared to KRAS wild-type in vivo (means ± S.E.M., n = 7 mice/group). e The tumor images of the indicated groups. f The tumor volume of the indicated groups. Tumors were measured every two days. g The tumor weight of the indicated groups. h Knocking down JOSD2 downregulated intratumor KRAS levels of KRAS-mutant xenografts are stronger than those of KRAS wild-type xenografts. The proteins isolated from the tumor were subjected to WB analysis. The significance analysis of (b, d, f, g) was conducted by One-way ANOVA (Bonferroni method was utilized to correct for multiple comparisons). The samples derived from the same experiment and processed on different gels in parallel are shown below: h one gel for JOSD2/GAPDH and another for KRAS. Source data are provided as a Source Data file.
