Research Paper Volume 16, Issue 16 pp 11776—11795

Proteomic and secretomic comparison of young and aged dermal fibroblasts highlights cytoskeleton as a key component during aging

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Figure 5. FLNB and ACTC1 analysis. (A) mRNA FLNB expression (RT-qPCR) of fibroblasts in quiescence or after 24h of TGF-β-1 stimulation (Mean±SEM, n= 6 for young and old cells); (B) mRNA expression quantified by RT-qPCR after transfection with specific siRNA for FLNB (siFLNB) or with Scramble siRNA (siScramble), in quiescence or after a 24h-TGF-β-1 stimulation (Mean±SD, n=6 per type); (C) Collagen gel contraction (lattice) of old fibroblasts either from control cultures or after transfection with specific siFLNB or with siScramble during 120h (Mean±SD, n=6); (D) Wound closure over 96h as measured by % of cell confluency for analysis of migration of old control fibroblasts (CT) or after transfection with FLNB or siScramble (Mean±SD, n=6); (E) mRNA ACTC1 expression (RT-qPCR) of fibroblasts in quiescence or after 24h of TGF-β-1 stimulation (Mean±SEM, n= 6 for young and old cells); (F) mRNA expression quantified by RT-qPCR after transfection with specific siRNA for ACTC1 (siACTC1) or with Scramble siRNA (siScramble), in quiescence or after a 24h-TGF-β-1 stimulation (Mean±SD, n=6 per type); (G) Wound closure over 96h as measured by % of cell confluency for analysis of migration of old control fibroblasts (CT) or after transfection with siACTC1 or siScramble (Mean±SD, n=6); (H) Collagen gel contraction (lattice) of young fibroblasts either from control cultures or after transfection with specific siACTC1 or with siScramble during 120h (Mean±SD, n=6); p-value *<0.05, **<0.01, ***<0.001.