ZMIZ1^△N185 expression transforms immortalized keratinocytes.

HaCaT cells were transfected with a Myc-tagged cDNA for full length or the truncated ZMIZ1^△N185 isoform (A) and protein expression was confirmed using a Myc antibody. (B) Colony-formation assays showed that HaCaT cells with ZMIZ1^△N185 expression form a greater number of colonies (top inset) and larger colonies (bottom inset) relative to HaCaT cells expressing the myc-tagged full length ZMIZ1 or empty vector control. Depletion of ZMIZ1 and ZMIZ2 in cuSCC cell lines decreases cell proliferation. Stable shRNA-mediated depletion of ZMIZ1 or ZMIZ22 in human cuSCC cell lines (D-E) A431 or (F-G) COLO16 confirmed by TaqMan analysis relative to cells transduced with a non-targeting shRNA (NTC). ZMIZ1 or ZMIZ2 depletion resulted in significantly decreased cellular proliferation in (E) A431 cells (overall P<0.0001; shNTC vs. shZMIZ1 q<0.0001; shNTC vs. shZMIZ2 q = 0.003; shZMIZ1 vs. shZMIZ2 q = 0.0023). (F) COLO16 (overall P = 0.0005; shNTC vs. shZMIZ1 q<0.0001; shNTC vs. shZMIZ2 q<0.0001; shZMIZ1 vs. shZMIZ2 q = 0.18). Statistical significance measured by 2-factor ANOVA followed by multiple pairwise comparisons and FDR-adjusted.