Fig. 6: Increased RAB5A-expression, cGAS perinuclear accumulation and DNA damage in human invasive ductal carcinoma.
From: Tissue fluidification promotes a cGAS–STING cytosolic DNA response in invasive breast cancer
a, Immunofluorescence images of control (CTR) and RAB5A MCF10.DCIS.com cells injected into mammary fat pads of immunocompromised mice. After one week, mice were fed doxycycline to induce RAB5A expression. Four weeks after doxycycline treatment, primary tumours were co-stained with DAPI and γH2AX, or DAPI and RAB5A. Scale bar, 100 μm. b, Percentage of γH2AX-positive cells per FOV. Data are mean ± s.d. (n = 2 experiments, with five mice per experiment), unpaired two-tailed t-test with Welch’s correction. c, Immunohistochemical analysis of cGAS in control (CTR) and RAB5A MCF10.DCIS.com cells injected into mammary fat pads of immunocompromised mice and treated as in a. Primary tumours were stained with an anti-cGAS antibody. Right panel: a higher magnification of the boxed region to highlight cGAS accumulation in perinuclear dots (black arrowheads) or crescent-like distribution (blue arrowhead). Scale bar 80 μm. d, The percentage of cells with perinuclear cGAS enrichment is expressed as the mean ± s.d. (each dot represents an FOV out of >5 FOVs per experimental condition in n = 2 independent experiments), two-tailed Mann–Whitney non-parametric test. e, Multiplex immunohistochemistry/Immunofluorescence (mIHC/IF) of RAB5A, γH2AX and DAPI in human DCIS. Magnified images from the selected yellow boxes are shown. Scale bar, 150 μm. f, The percentage of γH2AX-positive nuclei in cells that express high (>2 on a scale from 0,1,2,3) or low (<2 on a scale from 0,1,2,3) levels of RAB5A in the tumour bulk or margin (see Methods for details and Supplementary Video 13). The areas of n = 9 independent DCIS were analysed with more than 200 nuclei per area, two-tailed Mann–Whitney non-parametric test. g, Multiplex immunohistochemistry/immunofluorescence (mIHC/IF) of RAB5A, cGAS, and Dapi in human DCIS. Magnified images from the selected yellow boxes are shown. Scale bar, 300 μm. h, The percentage of cGAS-positive cells that express high (>2 on a scale from 0,1,2,3) or low (<2 on a scale from 0,1,2,3) levels of RAB5A in the tumour bulk, or the margin or the whole tumour (Methods). The areas of n = 9 independent DCIS were analysed with more than 200 nuclei per area, two-tailed Mann–Whitney non-parametric test and unpaired two-tailed t-test with Welch’s correction.
