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. 2023 May 22;34(8):1381–1397. doi: 10.1681/ASN.0000000000000154

Figure 5.

Figure 5

Anti–miR-147 protects against cell death and mitochondria dysfunction after ATP depletion in renal tubular cells. RPTCs were transfected with 100 nM anti–miR-147 or NC oligo and then subjected to 3 hours of CCCP treatment for ATP depletion with 2 hours of recovery (CCCP-R) or control incubation. (A) Representative images of cell morphology, 4′,6-diamidino-2-phenylindole staining of cell nuclei, PI staining, JC-1 aggregate, and JC-1 monomer staining. Scale bar=100 μm. (B) qPCR analysis of miR-147. (C) Percentage of cell death assessed morphologically. (D) Quantitative analysis of JC-1 green to red signal ratio. (E) Cellular ATP. (F) Immunoblots of cleaved caspase-3 with β-actin as the internal control. (G) Densitometry of cleaved caspase-3. (H) Measurement of OCR using an XF24 Extracellular Flux Analyzer. Quantitative data are expressed as mean±SD (n=4–10). *P < 0.05 versus control; #P < 0.05 versus CCCP-R with NC group. miR-147, microRNA-147; NC, negative control; OCR, oxygen consumption rate; PI, phosphatidylinositol; RPTCs, renal proximal tubular cells.