Extended Data Fig. 5: Response of MIRAS mutation-corrected fibroblast-like cells to HSV-1 infection, and control and patient fibroblasts response to TBEV or SARS-CoV-2 infection.
From: Ancestral allele of DNA polymerase gamma modifies antiviral tolerance
(a) Schematic for generation of induced fibroblast-like cells from iPSC cell lines. Created using BioRender.com. See method section for details. (b) POLG1 protein amount in controls, MIRAS patient and MIRAS mutation-corrected induced fibroblast-like cells. Western blot and quantification, loading control SDHA. Mean ± SEM. N = 4 controls (2 males, 2 females), 1 male patient, 2 independent MIRAS mutation-corrected clones of the male patient (Cr1, Cr2). (c) Viral protein, host inflammatory and necroptotic protein in cells (as in b) at 48 h post HSV-1 infection. Western blot and quantification, loading control SDHA. Mean ± SEM. (d) MtDNA amount at uninfected basal condition and 48 h post HSV-1 infection in cells (as in b). mtDNA, MT-12S to nuclear gene, B2M. Mean ± SEM. (e-h) Immune signalling (e,f) and necroptotic activation (g,h) of fibroblasts post 48 h of TBEV (e,g) or SARS-CoV-2 (f,h) infection. Western blot and quantification, loading control HSP60, arrowhead: band of interest. N = 6 female controls and patients per condition. Refer to Fig. 2g, h for other protein quantification. (i,j) Cellular viability of fibroblasts post 48 h of TBEV (i) and SARS-CoV-2 (j) infection. N = 6 female controls and patients per condition. In e, f, i and j, box plots show minimum-25th-50th(median)-75th percentile-maximum (whiskers extend to the smallest and largest value); two-tailed unpaired student’s t-test. Abbreviations: Pt, patient.
