Figure 6. ZBP1 regulates UVB-induced type I and III IFN responses in human keratinocytes.

A-B. Knockdown of ZBP1 in N/TERTs was performed using a lentivirus expressing either shRNA targeting Human ZBP1 or shcontrol. Baseline, UV, and UV+IFNα induced gene expression were assessed for (A.) Type I and III IFN (6h after UVB) and (B.) CCL5, a known ISG (24h after UVB); n=3 C. Generation of GFP/3XFLAG-tagged N/TERTs with overexpression of ZBP1 compared to GFP-tag only. D. IFN genes and ISGs were assessed by RT-qPCR at baseline in ZBP1 overexpressing cells (ZBP1 OE) or GFP-expressing control N/TERTs (GFP). E. Representative greyscale and merged images of confocal microscopy from N/TERTs with GFP or GFP-ZBP1 N/TERTs for cGAS (red) and DAPI (blue). Scale bar 20μm. F. Quantification of cytosolic cGAS using ratio of cGAS MFI in the cytosol versus nuclear cGAS MFI using CellProfiler. G. Representative confocal images of GFP-tagged N/TERTs or ZBP1 overexpressing N/TERTs of TOMM20 and Z-DNA at baseline and 3h after UVB exposure. Scale bar 10μm. Ordinary one –way ANOVA followed by Sidak’s multiple comparison test. Mean and SEM. *P<0.05, **P<0.01, ***P<0,001, ****P<0.0001.