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. 2021 Apr 23;118(5):1276–1288. doi: 10.1093/cvr/cvab149

Figure 7.

Figure 7

Cardiomyocyte-specific EGFR downregulation decreases expression of the PP2A B” regulatory subunit PR72. (A) RT-qPCR was used to measure the expression of Ppp2r3a in the LV of EGFRf/f (Cre−) vs. CM-EGFR-KD mice at 7, 9, and 16 weeks of age (n = 6 each). ***P < 0.001, two-tailed unpaired t-test. (B) Immunoblot analysis of PR72 expression in adult EGFRf/f (Cre−) vs. CM-EGFR-KD cardiac lysates, as summarized in histogram. Blots shown are representative of n = 11 cardiac lysates per genotype. ***P < 0.001, two-tailed unpaired t-test. (C) Immunoblot analysis of PR72 expression in adult CM-EGFR-KD mouse cardiomyocytes infected for 24 h with adenoviruses encoding lacZ or PR72, as summarized in histogram. Blots shown are representative of n = 4 independent cardiomyocyte preparations. **P < 0.01, two-tailed unpaired t-test. (D) Time to 50% relaxation measured in lacZ vs. PR72-infected AMCM from CM-EGFR-KD mice. *P < 0.05, two-tailed unpaired t-test. n = 14–15 individual cardiomyocytes isolated from n = 5 mice per adenovirus infection. (E) Time to 50% Ca2+ clearance in lacZ vs. PR72-infected AMCM from CM-EGFR-KD mice. *P < 0.05, two-tailed unpaired t-test. n = 16–19 individual cardiomyocytes isolated from n = 3 mice per condition. (F) Immunoblot analysis of PR72 expression in adult CM-EGFR-KD cardiac lysates, 2 weeks after infection with AAV9-lacZ or AAV9-PR72, as summarized in histogram. Blots shown are representative of n = 6 cardiac lysates per condition. *P < 0.05, two-tailed unpaired t-test. %EF (G) and %FS (H) of adult CM-EGFR-KD mice treated with AAV9-lacZ (n = 10, reference group from Figure 1E and F, respectively) vs. AAV9-PR72 (n = 5) were monitored via echocardiography for 4 weeks following AAV administration (at time 0). *P < 0.05, **P < 0.01, ***P < 0.001 vs. reference group CM-EGFR-KD+AAV9-lacZ at corresponding timepoints, ordinary two-way ANOVA with Bonferonni’s multiple comparisons test.