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Figure 2. Ischemic injury-induced JOSD2 expression in cardiomyocytes and post-MI hearts. (A) RT-qPCR analysis of Nppa (encoding ANP) and Josd2 mRNA levels in NRVMs under control conditions and after OGD for 6 h or 10 h (n = 4); (B) Western blots showing ANP and JOSD2 protein levels in NRVMs after OGD for the indicated durations; (C) Densitometric quantification of ANP and JOSD2 immunoblots (n = 3); (D) RT-qPCR analysis of Acta2 (encoding α-SMA) and Josd2 expression in murine hearts at 7 days post-MI, comparing sham, remote zone, and border zone (n = 5); (E) Western blots showing α-SMA and JOSD2 protein abundance in sham hearts and in the remote zone and border zone of MI hearts at day 7; (F) Densitometric quantification of α-SMA and JOSD2 (n = 4 per group); (G) Western blots of JOSD2 in sham hearts and at 12 h, 24 h, and 1 week after MI (n = 3); (H) Densitometric quantification of JOSD2 (n = 3); (I) Representative immunofluorescence images of JOSD2, cTnT, and DAPI staining in sham and 1-week post-MI hearts. Scale bar, 50 μm. All results are representative of at least three independent experiments. Data are presented as mean ± SD, and statistical analysis in (A), (C), (D), (F), and (H) was performed using one-way ANOVA with Tukey’s post hoc test. ns, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. JOSD2: Josephin domain-containing protein 2; RT-qPCR: reverse transcription quantitative polymerase chain reaction; MI: myocardial infarction; OGD: oxygen-glucose deprivation; ANP: atrial natriuretic peptide; NRVMs: neonatal rat ventricular myocytes; α-SMA: α-smooth muscle actin; ANOVA: analysis of variance; SD: standard deviation; cTnT: cardiac troponin T; WT: wild-type; DAPI: :4’,6-diamidino-2-phenylindole.