Figure 2.

Circ-Calm4 inhibits pyroptosis in a mouse model of hypoxia-induced pulmonary hypertension (PH). A and B, Knockdown of circ-Calm4 by sh-circ-Calm4 countered the hypoxia-induced upregulation of Caspase-1, NLRP3 (nucleotide-binding oligomerization segment-like receptor family 3), ASC (apoptosis-associated speck-like protein containing a caspase recruitment segment ),IL-18 (interleukin-18), and IL-1β (interleukin-1β) at the protein and mRNA levels (n=6; in Western blot, the image of NLPR3 and Caspase-1 used the same actin). C and D, Knockdown of circ-Calm4 by sh-circ-Calm4 reversed the hypoxia-induced upregulation of Caspase-1 and IL-18 in mouse lung tissues. Scale bars=100 µm. Lung sections stained with Caspase-1 (green) and IL-18 (green), pulmonary smooth muscle stained with α-SMA (red), and DAPI for nuclear staining. Each datapoint in the figure represents a unique biological replicate. The data are presented as the means±SD. Statistical analysis was performed with 1-way ANOVA followed by Bonferroni correction. The graph B-ASC was analyzed by the Kruskal-Wallis test followed by Dunn post-test. *P<0.05, **P<0.01, ***P<0.001.