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. 2022 May 21;28:920–934. doi: 10.1016/j.omtn.2022.05.033

Figure 6.

Figure 6

RPS4XL inhibits HSC70 glycosylation, which inhibits hypoxia-induced pyroptosis in PASMCs

(A) Schematic diagram of the in vitro purification of the HSC70 protein domains. Microscale thermophoresis of the HSC70 1–393 aa and different concentrations of RPS4XL. (B) Bioinformatics analysis the N-glycosylation site in the 1–393 aa domain of HSC70, and schematic diagram of the HSC70 glycosylation site mutation. (C) LDH release assay in hypoxic and normoxic PASMCs transfected with OE-HSC70, Mut-HSC70, or treated with 10 μg/mL RPS4XL. (D) PI staining in hypoxic and normoxic PASMCs transfected with OE-HSC70, Mut-HSC70, or treated with 10 μg/mL RPS4XL (scale bar, 50 μm). Western blotting analysis of (E) c-caspase-1, (F) NLRP3, ASC, IL-1β, and IL-18 in hypoxic and normoxic PASMCs transfected with OE-HSC70, Mut-HSC70, or treated with 10 μg/mL RPS4XL. All values are represented as the mean ± SEM (∗p < 0.05, and ∗∗p < 0.01; n ≥ 3). NOR, normoxia; HYP, hypoxia.