Fig. 2: Aberrant immune response of fibroblasts of patients with MIRAS to bona fide viral infection.
From: Ancestral allele of DNA polymerase gamma modifies antiviral tolerance
a, Schematic of HSV-1 infection of human primary fibroblasts. b,c, Viral load and host inflammatory protein level modulation in fibroblasts during HSV-1 infection. Western blot (b) and quantification (c) of protein amounts over time during infection in a control and patient fibroblast line. Protein loading stain was used as the loading control. Exp., exposure. d, Quantification of viral and host cellular protein amounts at the specific timepoint after HSV-1 infection. n = 6 female controls and patients. The western blot is shown in Extended Data Fig. 4a–c,e. The loading control was HSP60. e, mtDNA levels in fibroblasts (as in d) during HSV-1 infection. qPCR analysis of mtDNA (MT-12S (MT-RNR1)) relative to a nuclear gene (B2M). f, The viral load in fibroblasts after 48 h of TBEV infection. Immunofluorescence analysis of TBEV antigen (green) and dsRNA (red; detects viral RNA) with DAPI co-staining (blue) is shown. Each channel is shown in greyscale. Scale bars, 100 μm. n = 2 female control individuals and patients. g, Quantification of the viral load in fibroblasts after 48 h of TBEV or SARS-CoV-2 infection. Top, the percentage of TBEV-positive cells. n = 4 control and 3 MIRAS images of 2 female control individuals and patients shown in f. Bottom, quantification of the western blot analysis of SARS-CoV-2 nucleocapsid protein (Extended Data Fig. 5h). The loading control was HSP60. n = 6 female control individuals and patients. h, POLG1 and inflammatory protein level in fibroblasts after 48 h of TBEV or SARS-CoV-2 infection. Quantification of the western blot is shown (Extended Data Fig. 5e–h). The loading control was HSP60. n = 6 female control individuals and patients. i, Schematic of the response of cells from patients with MIRAS to virus infection. For d, e, g (bottom) and h, the box plots show minimum to maximum values (whiskers), 25th to 75th percentiles (box limits) and median (centre line). For g (top), data are mean ± s.e.m. Statistical analysis was performed using two-tailed unpaired Student’s t-tests. See also Extended Data Figs. 4 and 5.
