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Fig. 4 Cu2+ directly suppressed smyd1b and myofibrillogenesis independent of the integral function of cox17 or atp7b. (A1) Cu2+ directly suppressed the transcriptional activity of the smyd1b promoter and thus the transcriptional activation of Myog and Srfa. (A2) WB assays of the ectopic expression of Myc tagged Myog and Srfa protein, respectively, with Myc tagged hEaf2 as the positive control.27, 28 B, WISH assays of mylpfa (B1-B6), smyhc1l (B8-B13), and smyd1b (B15-B20) expression in cox17−/− or atp7b−/− mutants with or without Cu2+ stresses. B7, B14, B21, the calculated expression of mylpfa, smyhc1l, or smyd1b in different groups. Each dot represents the signal level in a representative individual embryo. Each experiment was repeated two or three times with similar results, and a representative result is shown. B1-B6, B8-B13, B15-B20, lateral view, and anterior to the left. B1-B6, B8-B13, B15-B20, scale bar, 250 μm; *P < .05, **P < .01, ***P < .001. NS, not significant