An immunoprecipitation kit (Proteintech, Wuhan, China) was utilized to evaluate the interactions of p22phox with p47phox and of p22phox with NOX4 in KGN cells according to the manufacturer's instructions. In brief, cell lysates were preincubated with anti-mouse p22phox (sc-271968, Santa Cruz Biotechnology) or anti-mouse p47phox antibodies (sc-17844, Santa Cruz Biotechnology) at 4°C overnight and incubated with Protein A Sepharose beads at 4°C for 4 h. Protein complexes were obtained after washed and eluted 5 times, and then detected by western blotting using anti-mouse p22phox (sc-271968, Santa Cruz Biotechnology), anti-mouse p47phox (sc-17844, Santa Cruz Biotechnology), and anti-rabbit NOX4 (ab109225, Abcam) antibodies.
Zhou X.Y., Zhang J., Li Y., Chen Y.X., Wu X.M., Li X., Zhang X.F., Ma L.Z., Yang Y.Z., Zheng K.M., Liu Y.D., Wang Z, & Chen S.L. (2021). Advanced Oxidation Protein Products Induce G1/G0-Phase Arrest in Ovarian Granulosa Cells via the ROS-JNK/p38 MAPK-p21 Pathway in Premature Ovarian Insufficiency. Oxidative Medicine and Cellular Longevity, 2021, 6634718.
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