Fig. 6: Inhibition of protein synthesis restores nuclear envelope abnormalities in NGPS cells.
From: A multiparametric anti-aging CRISPR screen uncovers a role for BAF in protein synthesis regulation
A Nascent protein synthesis assay based on fluorescence intensity of HPG AF 488. NGPS2 or NGPS2 corrected cells (NGPS WT) were imaged with the high throughput microscope following siRNA depletion of 41 of the validated screen hits. Depletion of genes highlighted in blue show significant (one-way ANOVA with Dunnett’s multiple comparisons test, p < 0.05) reduction of HPG incorporation compared to NGPS2 cells transfected with a non-targeting siRNA (red dotted line). Shown are averages ± SD of 3 independent experiments in 500 cells. B Quantification of emerin nuclear intensity in NGPS1 and NGPS2 cell lines compared to WT, upon protein synthesis inhibition using cycloheximide (CHX) or silvestrol. Superplots of the 3 independent experiments are shown, and nested one-way ANOVA was used for statistical comparisons with Dunnett’s multiple comparison testing. C Representative immunofluorescence images of emerin staining upon protein synthesis inhibition by treatment with CHX or silvestrol.
