Fig. 5

Rsrc1-161aa regulates mitochondrial ribosome assembly and protein translation. a RT-qPCR detection of mitochondrial ribosomal gene transcript levels in wild-type and Rsrc1^+/+/circRsrc1^−/− cells. RNA data were normalized by 18S (n = 3). b Mass spectrometry targeted quantitative detection of proteins encoded by mitochondria in wild-type, Rsrc1^+/+/circRsrc1^−/− and rescue cells (n = 3). c Western blotting of mitochondrial respiratory proteins in wild-type, Rsrc1^+/+/circRsrc1^−/− and rescue cells. The statistical graph shows the gray values after three repetitions (n = 3). d Flow cytometry was used to separate spermatocytes from testis, and Western blotting was performed to detect mitochondrial respiratory proteins. The statistical graph shown the gray values after three repetitions. e Western blotting of mitochondrial respiratory protein in mouse sperm. The statistical graph shows the gray values after three repetitions (n = 3). f RIP-qPCR experiment in wild-type and Rsrc1^+/+/circRsrc1^−/− cells using the anti-C1qbp antibody (n = 3). g Cell lysates from wild-type, Rsrc1^+/+/circRsrc1^−/.⁻ and rescue cells were separated by 15 ~ 30% sucrose density gradient and analyzed using antibodies against Mrps22 (28S small subunit) and Mrpl28 (39S large subunit) proteins. All data are means ± SEM. When performing statistical tests between two groups of data, tested using a two-tailed unpaired t-test. When performing statistical tests between three or more groups of data, tested using one-way ANOVA with Tukey’s post hoc analysis. *P < 0.05; **P < 0.01; ***P < 0.001