Figure 5.
The rescue of phenotypic changes in circSlc8a1 and cA-circSlc8a1 double transgenic mice
(A) The levels of unbound circSlc8a1 were detected by gene-specific RT-qPCR in circSlc8a1(+), cA-circSlc8a1(+), double transgenic mice, and the litter-matched negative mice. n = 10; ∗∗p < 0.01. (B) The levels of unbound circSlc8a1 were detected by RNA pull-down assay with circSlc8a1 probes in circSlc8a1(+), cA-circSlc8a1(+), double transgenic mice, and the litter-matched negative mice. n = 10; ∗∗p < 0.01. (C) The body weight difference in circSlc8a1(+), cA-circSlc8a1(+), double transgenic mice compared with the average of the litter-matched negative mice. n = 10; ∗∗p < 0.01. (D) Representative photographs of the whole hearts and WGA staining showed that cardiac hypertrophy was not induced in the double transgenic mice compared with the cA-circSlc8a1(+)-transgenic mice. n = 10. (E) Representative photographs of H&E and Oil Red O staining showed that hepatic steatosis was not induced in the double transgenic mice compared with the cA-circSlc8a1(+)-transgenic mice. n = 10. (F) The levels of unbound circSlc8a1 were detected by gene-specific RT-qPCR after 0.25, 0.5, 1, 2, or 3 μg cA-circSlc8a1 plasmids per 1 × 106 cells or the vectors were transfected. n = 6; ∗∗p < 0.01 versus vector. (G) The levels of unbound circSlc8a1 were detected by the RNA pull-down assay with circSlc8a1 probes after 0.25, 0.5, 1, 2, or 3 μg cA-circSlc8a1 plasmids per 1 × 106 cells or the vectors were transfected. n = 6; ∗∗p < 0.01 versus vector. (H) The unbound circSlc8a1 levels after transfection of vector, cA-circSlc8a1 plasmids and the cA-circSlc8a1 plasmids with mutated introns (lin-cA-circSlc). The levels of unbound circSlc8a1 were detected by gene-specific RT-qPCR. n = 6; ∗∗p < 0.01 versus vector. (I) The unbound circSlc8a1 levels were detected by the RNA pull-down assay after transfection with the vector, cA-circSlc8a1 plasmids and the cA-circSlc8a1 plasmids with mutated introns (lin-cA-circSlc). n = 6; ∗∗p < 0.01 versus vector.