Fig. 6

TDRKH-AS1 induces pyroptosis by binding to PDIA4 in HUVECs. A The TDRKH-AS1-protein-pathway multilayer interaction network, containing TDRKH-AS1, 138 proteins interacting with TDRKH-AS1 and 3 enriched pathway of proteins involved in, 138 lncRNA-protein interactions, and 24 protein-pathway relationships. B Enriched pathways of proteins interacting with TDRKH-AS1. C The interaction between TDRKH-AS1 and PDIA4 using RNA pull-down was validated by western blotting. D Binding proteins of TDRKH-AS1 detected by RIP assays and qRT-PCR. E TDRKH-AS1 overexpression increases the expression of PDIA4 on mRNA and protein levels, while TDRKH-AS1 knockdown showed the opposite effect. F PDIA4 promoter contains four latent binding sites with TDRKH-AS1. (Primers for the regions marked by purple). G, H ChIRP was performed followed by qPCR to verified that TDRKH-AS1 bound to the promoter of PDIA4. The specific binding of TDRKH-AS1 with its probes was verified using qRT-PCR. I Knockdown PDIA4 partially alleviated the pyroptosis-associated proteins expressions trigged by overexpression TDRKH-AS1 in HUVECs using western blotting. J–L The release of LDH, IL-1β and IL-18 induced by overexpression of TDRKH-AS1 were partially inhibited via knockdown of PDIA4 in HUVECs using LDH activity assay and ELISA. The data are represented as the mean ± SD; **p < 0.01, *p < 0.05