Research Paper Volume 15, Issue 24 pp 14749—14763

Downregulating miR-184 relieves calcium oxalate crystal-mediated renal cell damage via activating the Rap1 signaling pathway

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Figure 2. miR-184 knockdown abated renal cell damage mediated by CaOx. CaOx (100 μg/mL) was used to treat HK2 and HPC cells, with or without miR-184-in transfection, for 24 hours. (A) qRT-PCR analysis of miR-184 expression. (B, C) Cell viability was examined by CCK8 assay. (D) TUNEL staining monitored cell apoptosis. Scale bar=100 μm. (E) The profiles of apoptosis-related proteins (Mcl1, Bcl-XL, Caspase-3) were determined by Western blotting. (F) The protein profiles of the NF-κB and Nrf2/HO-1 pathways were confirmed through Western blotting. (G) The protein levels of NF-κB p65 and Nrf2 in the nucleus were verified through Western blotting. (H, I) ELISA revealed the levels of the inflammatory factors IL-6, TNF-α, MCP1, and ICAM1. (J) Cell immunofluorescence was used to examine the level of ROS. Scale bar=100 μm. **P<0.01, ***P<0.001 (vs. the Con group). ^P<0.05, ^^P<0.01, ^^^P<0.001 (vs. CaOx+miR-in). N=3.