Fig. 1: NGPS fibroblasts show distinctive phenotypes to HGPS.
From: A multiparametric anti-aging CRISPR screen uncovers a role for BAF in protein synthesis regulation
A Representative transmission electron micrographs of WT, NGPS1 and NGPS2 fibroblasts (males). B Representative high resolution microscopy images showing the expression and localization of lamin B1 and emerin in WT and NGPS fibroblasts. C Immunofluorescence images showing DNA damage foci (53BP1) and nucleocytoplasmic transport (Ran) in WT and NGPS fibroblasts. Images were obtained with the CX7 high-content microscope and quantified in (D) and (E) using the HCS StudioTM software. D Superplot of the data (3 technical replicates in 3 independent experiments), lines indicate average values, statistical analysis using nested one-way ANOVA with Dunnett’s multiple comparisons. E Data from 3 independent experiments, 2-3 technical replicates each, lines indicate average values, statistical comparison using one-way ANOVA with Dunnett’s multiple comparisons. F High resolution microscopy images of the heterochromatin marker HP1γ in WT and NGPS cells. G Nuclear HP1γ levels quantified using the high-content microscope in 2 independent experiments, each averaging 500 nuclei in 3 wells. Means ± SD are shown. For the NGPS cells statistical testing used one-way ANOVA with Dunnett’s multiple comparisons; HGPS cells were compared to WT using two-tailed unpaired t-test. H Representative Western blot and (I) quantitative analysis of 4 independent experiments showing HP1γ in NGPS cells compared to WT. J–L Representative western blots and quantitative analysis of 3 independent experiments, showing the heterochromatin marks H3K9me3 (J) and H3K79me2 (K) or the aging and senescence marker p21 (L) in NGPS cells compared to WT. In I–L data points are overlaid on columns indicating the mean +/- SD, and statistical analysis used one-way ANOVA with Dunnett’s multiple comparisons.
