Validation of shRNA for MLKL in cultured DRG neurons and N2a cells. (A) Immunoblot analysis of MLKL from DRG neurons expressing control shRNA (shCon) or MLKL targeted shRNA. Arrow (MLKL band), arrowhead (nonspecific band). TUJ1 is used as a loading control. (B) Quantification of MLKL knockdown in DRG neurons expressing shMLKL as in A. Data represent the mean ± SEM, n = 5; two-tailed unpaired t test, P < 0.0001; *, P < 0.05; **, P < 0.01; and ***, P < 0.001. (C) Immunoblot analysis of MLKL from N2a cells expressing control or MLKL targeted shRNA. TUJ1 is used as a loading control.