Fig. 2: Knockdown of DAPK1 confers increased sensitivity to necroptosis on HT-29 cells.
From: Tumor suppressor death-associated protein kinase 1 inhibits necroptosis by p38 MAPK activation
a Knockdown of DAPK1 in HT-29 cells. HT-29 cells were transduced with pLL3.7-shCtrl or pLL3.7-shDAPK1, sorted, and then expressions of DAPK1, RIPK1 and RIPK3 were determined. b Increased zVAD+BV6-induced necroptosis in DAPK1-deficient HT-29 cells. Control and DAPK1-knockdown HT-29 cells were treated with zVAD (20 μM), BV6 (1 μM), Nec-1 (40 μM) as indicated for 24 h, cell death was quantitated by PI staining. Values are mean ± SD of triplicates in a single experiment. ***P < 0.001 for unpaired t-test. Results have been confirmed in three independent experiments. c Enhanced zVAD+BV6-induced phosphorylation of RIPK1, RIPK3 and MLKL in DAPK1-deficient HT-29 cells. Control and DAPK1-knockdown HT-29 cells were treated with zVAD+BV6 (0.5 μM, Z + B) and contents of RIPK1, pS166-RIPK1, RIPK3, pS227-RIPK3, MLKL, and pMLKL in cell lysates were determined at the indicated time points. Right panel, quantitation of pRIPK1(S166), pRIPK3 and pMLKL from three independent experiments using normalized intensity of pRIPK1(S166), pRIPK3 and pMLKL in DAPK1-knockdown HT29 cells at 9 h as 1. **P < 0.01, ***P < 0.001 for two-way ANOVA followed by a Tukey’s multiple comparison test.
