Figure 8.

Inhibition of AT1R and AT2R mitigates the expression of Fas/FasL signaling molecules in Ang II-induced HK-2 cells. Effects of FasL blockade via neutralizing human Fas ligand/TNFSF6 antibody on Ang II-induced necroptosis in HK-2 cells were determined. HK-2 cells were pretreated with 3 µg/ml neutralizing human Fas ligand/TNFSF6 antibody for 2 h and exposed to 10⁻⁹ M Ang II for 24 h. Then, Western blotting was performed to detect Fas and FasL (A,D,E), RIP3, MLKL, p-RIP3, and p-MLKL (B,F–I) levels and β-actin was used as a loading control. C shows representative images of immunofluorescence staining for RIP3 (red fluorescence) and in situ fluorescence TUNEL staining (green fluorescence). Scale bars represent 50 μm. (J) The data are presented as the % ratio of necroptotic HK-2 cells (TUNEL-positive and RIP3-positive cells). The results shown are representative of three independent experiments. The intensities of the bands were determined quantitatively using Image-Pro Plus 6.0 **p < 0.01 versus control; ^##p < 0.01 versus the Ang II-induced group. Ang II: angiotensin II.