Journal: Cell Death & Disease

Article Title: Regulator of calcineurin 1 deletion attenuates mitochondrial dysfunction and apoptosis in acute kidney injury through JNK/Mff signaling pathway

doi: 10.1038/s41419-022-05220-x

Figure Lengend Snippet: A siRNA against JNK and si-NC were transfected into cells. The expression level of JNK was monitored via Western blotting. ( n = 5) B An IF assay for p-JNK, and DAPI was used to tag the nucleus. Scale bar, 10 μm. n = 3. C The renal tubular epithelial cell line HK-2 was used with HR, and siRNA against JNK and si-NC were transfected into cells. The expression levels of Pro-caspase-3, Cle-caspase-3, Pro-caspase-9, Cle-caspase-9, Bax, and Bcl-2 were detected by Western blotting ( n = 5). D Mitochondrial morphology of HK-2 was assessed by MitoTracker TM Deep Red staining, and the average length of mitochondria was measured. Scale bar, 10 μm n = 3. E Western blotting was performed to analyze the expression of p-Drp1 S616 , Drp1, p-Mff, Mff, Fis1, Mfn1, Mfn2, and Opa1 in HK-2 cells ( n = 5). F Mitochondrial function was measured with an ATP assay kit ( n = 4). G – I Representative Co-IP analysis of RCAN1, JNK, and Mff in HK-2 cells under HR injury ( n = 3). J – L The co-localization of RCAN1 and p-JNK, Mff, and JNK, as well as Mff and RCAN1, were detected by IF, and DAPI was used to tag the nucleus. Scale bar, 10 μm. n = 3. * p < 0.05, ** p < 0.01.

Article Snippet: Nuclei were visualized by staining with DAPI (G1012, Servicebio) for 10 min at room temperature.

Techniques: Transfection, Expressing, Western Blot, Staining, ATP Assay, Co-Immunoprecipitation Assay