Fig. 8: Pharmacological inhibition of RIPK2 inactivates RIPK2/MKK7/c-Myc signaling and suppresses PC metastatic outgrowth.

a Representative immunoblots (left) and quantification (right) of p-MKK7-S271 in total lysates of RIPK2-KO HEK293T cells under the indicated conditions (2 h treatment in the presence of 10% FBS) (n = 3 biologically independent samples per group). b, c Representative immunoblots of the indicated proteins in total lysates of b RIPK2-KO HEK293T cells or c 22Rv1 cells under the indicated conditions (2 h treatment). Experiments were repeated three times independently with similar results (b, c). d Quantification of 3D spheroid invasion assays of PC3 cells treated with vehicle control, 10 µM GSK583, or 5 µM ponatinib (n = 6 biologically independent samples per group). e Bar plot of anchorage-dependent colony formation of PC3, DU145, and 22Rv1 cells treated with vehicle control or 10 µM GSK583 (n = 6 biologically independent samples per group). f Drug response curve of anchorage-dependent colony formation of PC3 cells treated with vehicle control or the indicated concentrations of ponatinib (n = 6 biologically independent samples per group). g Schematic diagram of drug treatment for the inhibition of PC metastasis in vivo. Created with BioRender.com. h Effect of GSK583 treatment (10 mg/kg/day) on total BLI intensities in biologically independent mice (vehicle, n = 5; GSK583, n = 7), starting on day 9 following the i.c. injection. i Representative BLI images of the ventral (upper) and dorsal (lower) sides of mice treated with vehicle control or GSK583 for 4 weeks. j Effect of low-dose (6 mg/kg/day) or high dose (30 mg/kg/day for 10 days followed by 15 mg/kg/day for 18 days) ponatinib treatment on total BLI intensities in biologically independent mice (vehicle, n = 6; low-dose ponatinib, n = 8; high-dose ponatinib, n = 7), starting on day 9 following the i.c. injection. Pon: ponatinib. k Representative BLI images of the ventral (upper) and dorsal (lower) sides of mice treated with vehicle control, low-dose ponatinib, or high-dose ponatinib for 4 weeks. Nominal p-values were determined by unpaired two-tailed Student’s t-test (a, d–f) or two-way ANOVA (h, j). Data are Mean ± SEM (a, d–f, h, j).