Fig. 4.

CircSSR1 regulated pyroptosis and endoplasmic stress through acting on SSR1 expression under hypoxia. A and B, qRT-PCR and western blot showed over-expression of circSSR1 had no significance on mRNA (n = 6) but affect SSR1 translation (n = 4). C, DAVID website demonstrated Gene Ontology (GO) analysis showed SSR1 was related with unfold protein response. D, Endoplasmic reticulum specific probe (Green) and SSR1 (Red) can localization proved by ER-Tracker. E, Interfere with SSR1 under hypoxia can prevent hypoxia-induced LDH release (n = 6). F, Transfection siRNA decrease immunofluorescence intensity of IL-1β in HYP. G, Pyroptosis markable proteins expression after transfection siSSR1 (n = 5). H, PI staining showed the positive cells after transfect siRNA (n = 6). I, Western blot showed ER stress marker proteins expression after transfected with siSSR1 (n = 5–6). J, Immunofluorescence intensity of GRP78 decreased after transfect siRNA (n = 6). Scale bars, 50 μm. All values are presented as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001. NOR, normoxia and HYP, hypoxia. NC: negative control