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. 2023 Jan 12;14(1):19. doi: 10.1038/s41419-023-05553-1

Fig. 4. Knock-down of HS2/HS3 attenuates senescence of human lung fibroblasts.

Fig. 4

qRT-PCR analysis (A) or immune DNA-RNA FISH (B) of HS2/HS3 (red) and αSMA (green) in HFL1 transfected with scrambled siRNA or siRNA targeting human HS2/HS3 were serum starved and left untreated or treated for 4 d with 30 μg/ml bleomycin. Mean/SD from three independent experiments is shown. C HFL1 cells were transfected with siRNA, serum-starved for 6 days and stained with rhodamine-phalloidin (Rh-Ph). One out of five representative images is shown. D Brightfield image of HFL1 cells 6 d after transfection with siRNAs, serum starved untreated or treated with bleomycin. Cell number of cells grown for 4 d at 0.5% FCS media is presented at the graph (right); n = 3, 3 independent experiments. E xCELLigence real-time cell analysis (RTCA) dual purpose (DP) system (Agilent, USA) -recorded growth of primary human adult lung fibroblasts transfected with si-scr or si-HS2/HS3 and treated with bleomycin. F SA-βgal staining of starved HFL1 cells treated for 4 d with TGFβ, conditioned media from A549 tumor cells or 30 μg/ml bleomycin. One out of five representative images for each treatment is shown. Percentage of SA-βgal-positive cells per image has been calculated from 5 independent images and shown in the graph. Scale bars are shown in the images.