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. Author manuscript; available in PMC: 2021 Mar 1.
Published in final edited form as: Proteomics. 2019 Dec 30;20(5-6):e1800406. doi: 10.1002/pmic.201800406

Figure 2. Progerin causes downregulation of RAD51.

Figure 2.

(A) Re-analysis of published RNAseq data performed in 5 lines of normal fibroblasts (NF) and 3 lines of HGPS patients-derived fibroblasts growing in culture for 90 days [7] (accession # GSE97986). Expression of DNA repair genes shows a decrease in the BRCA pathway. *p represents p-value of statistical significance and ns=no significant (p>0.05). (B) HDF-GFP-Progerin cells were treated with Doxy for increasing days and the levels of RAD51, P-RPAS33, VDR, and proteins in the cGAS/STING/IFN pathway monitored by immunoblotting. β-tubulin is the loading control. (C) Immunofluorescence in HDF-GFP-progerin cells treated with Doxy for 4 days. Graphs show quantitation of RAD51 (top) and P-RPAS33 (bottom) labeling intensity (relative fluorescence units). GFP staining was used to demarcate nuclei and intensity of RAD51 and P-RPAS33 labeling measured using ImageJ program. Results are the average ± s.e.m. of 3 independent experiments. (D) Relative expression of RAD51 transcripts by qRT-PCR in HDF-GFP-Progerin cells treated with Doxy for increasing days. Results are average ± s.e.m. of 2 biological repeats, each performed in triplicate.