Fig. 5: Disruption of SMYD3 and SHCBP1 decreases the activities of oncogenic pathways.

a, b Representative images of tumor slices from G600 tumors without (control) or with BCI-121 treatment at a concentration of 300 µM for 7 days (a). The MTT assay was performed after harvesting the slices 7 days later and the quantifications of cell viability (b) in (a) (n = 3 mice/group). The IHC against antibodies of Smyd3, H3K4me3, Ki67 (c) and quantifications of protein intensities (d) from the same cohort of tumor slices in (a, b) (n = 3 mice/group). Scale bar is 50 μm. Representative tumor images from nude mice implanted with G600 parental and sgSmyd3-G600 (e) and sgShcbp1-G600 (f) in mammary fat pads with 2 × 10⁵ cells per mouse for 27 days (n = 10-12 mice/group). The tumor weight plots from the same cohort of mice with expression of sgSmyd3 (g) or sgShcbp1 (h). The protein levels of Smyd3, Shcbp1, Kras, pMek, and pErk in tumors initiated with parental G600, sgSmyd3-G600 (i), and sgShcbp1-G600 cells (j) in nude mice as shown by Western blots. k The cell growth curve of MDA-MB-231 (231) parental, sgSMYD3-231, and sgSHCBP1-231 cells was measured by IncuCyte. Tumor images of parental 231, sgSMYD3-231, and sgSHCBP1-231 cells (l) and plot of tumor weight (m) in nude mice (n = 12 mice/group). n The plot of relative spleen weight from the nude mice with the implantation of parental MDA-MB-231 (231), sgSMYD3-231, and sgSHCBP1-231 cells at 2 × 10⁶ cells per mammary fat pad for 70 days (n = 12 mice/group). The protein levels of SMYD3, SHCBP1, KRAS, GRB2, pMEK, and pERK in tumors initiated with parental 231, sgSMYD3 (o), and sgSHCBP1 cells (p) in nude mice by Western blots. q Summary of tumor growth after disrupting either Smyd3 or Shcbp1 in mice.