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. 2023 Mar 28;132(9):1110–1126. doi: 10.1161/CIRCRESAHA.122.322227

Figure 2.

Figure 2.

ZBP1 (Z-DNA binding protein 1) knockdown exacerbates increases in inflammatory cytokines in cardiomyocytes treated with mitochondrial DNA (mtDNA). A, mRNA levels of Zbp1 in neonatal rat ventricular myocytes (NRVMs) treated with or without small interfering RNA (siRNA) for ZBP1 (10 nmol/L) in the presence or absence of mtDNA (1000 ng/mL) for 24 hours (n=15). The experiment was conducted 5×. B, Representative immunoblots of ZBP1 (n=9), RIPK3 (receptor interacting protein kinase; n=12), phosphorylated NF-κB (nuclear factor-κB; Ser 536), NF-κB (n=15), NLRP3 (nucleotide-binding domain and leucine-rich-repeat family pyrin domain containing 3; n=9), and GAPDH in NRVMs treated with or without small interfering RNA (siRNA) for ZBP1 (10 nmol/L) in the presence or absence of mtDNA (1000 ng/mL) for 24 hours. The experiment was conducted 3 (ZBP1 and NLRP3), 4 (RIPK3), and 5× (NF-κB). C, mRNA levels of Il1b (n=15) and Il6 (n=18) in NRVMs treated with or without siRNA for ZBP1 (10 nmol/L) in the presence or absence of mtDNA (1000 ng/mL) for 24 hours. The experiment was conducted 5 (Il1b) or 6 (Il6) ×. D, Representative immunoblots of TBK1, phosphorylated TBK1, and GAPDH in NRVMs treated with or without siRNA for ZBP1 (10 nmol/L) in the presence or absence of mtDNA (1000 ng/mL) for 24 hours (n=12). The experiment was conducted 4×. Error bars denote standard errors. Data were analyzed using the Wilcoxon rank sum test (C and E; adjust=3), and 2-way ANOVA followed by Tukey multiple comparisons test (A, B, and D).