Gasotransmitter CO Attenuates Bleomycin-Induced Fibroblast Senescence via Induction of Stress Granule Formation

Figure 5

ISRIB abrogates CO-mediated SG formation and recovers BLM-induced cellular senescence. (a) WI-38 cells were treated with 40 μM CORM-A1 in the presence or absence of ISRIB (200 nM) for 6 h. Immunofluorescence assay was performed to detect the formation of SGs by visualizing the colocalization of TIA-1(red) and G3BP1 (green). WI-38 cells were pretreated with CORM-A1 (40 μM) with or without ISRIB (200 nM) for 6 h followed by the challenge of bleomycin (25 μg/ml) for 96 h. Then, SA-β-gal staining (b) and γ-H2AX foci (c) were detected, and immunofluorescence assay (d) was performed to assess the coaggregates of TIA-1 (red) and PAI-1 (green). The mRNA expressions of p21 (e), IL-6 (f), TNF-α (g), and IL-1β (h) by qRT-PCR. The secretions of PAI-1 (i), IL-6 (j), TNF-α (k), and IL-1β (l) were detected by ELISA. Quantitative data are expressed as the ( determined in three independent experiments). , , and .

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