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. 2016 Sep 30;7(51):84608–84622. doi: 10.18632/oncotarget.12358

Figure 5. Isoform specific activation of p38MAPK is critical for HS06's effects on CSCs growth and self-renewal.

Figure 5

a. Immuno-precipitation (IP) with anti-pp38 followed by western blotting (WB) with isoform specific anti-p38 antibody and b. the corresponding bar graph representation of relative expression shows activation of α and β isoforms, inhibition of δ isoform, and no significant change in γ isoform upon HS06 treatment (100 μM @ 3hr) in HT29 cells. c. Effect of HS06's inhibition on HT29 CSC self-renewal (3° spheroids) is completely attenuated in the presence of p38α siRNA (KD) compared to scrambled controls. d. Western blot analysis and e. the corresponding bar graph representation of relative expression of CSC markers (CD133, CD44) and self-renewal factors (c-MYC, BMI-1) shows complete reversal HS06's inhibition in the presence of p38α siRNA at 24 hr. GAPDH was used as loading control. f. Effect of HS06 on HT29 CSCs’ self-renewal (3° spheroid formation) is enhanced in the presence of p38δ siRNA compared to scrambled controls. GAPDH was used as loading control. Error bars represent ±1 SEM. * p-value < 0.005 compared to vehicle control. # p-value < 0.005 compared to HS06.