shRNA suppresses CAPN5 in cell lines. Quantitative PCR and immunoblot results for HEK293T-CAPN5-p.R243L (A- C) and SH-SY5Y (D- F) show knockdown effect of shRNA transfection. A, D. CAPN5 mRNA levels relative to GAPDH (HEK293T-CAPN5-p.R243L) or β-actin (SH-SY5Y) mRNA after transfection with each shRNA vectors as shown by percent knockdown of the respective vectors compared to the negative control vector. Quantitative PCR data are from average of three independent experiments. B, E. Immunoblots of transfected cell lysates show calpain-5, with GAPDH immunostaining as the loading control. C, F. Immunoblot band densities depict the average of three independent experiments. Bar graphs were represented as a mean ± SEM.