Fig. 4.

NEDD4L regulates ferroptosis in ischemic stroke by controlling TFRC ubiquitination. A. The BioGRID database was used to predict the protein interaction network of NEDD4L. B. Ubibroswer database was used to predict TRFC-associated ubiquitinated E3 ligases. C. QRT-PCR and WB were used to detect the mRNA and protein expression levels of NEDD4L after MCAO treatment, n = 6. D. QRT-PCR or WB was used to detect the mRNA expression levels of NEDD4L after OGD/R or NEDD4L treatment, n = 3. E. CO-IP was used to verify the binding of NEDD4L and TFRC in HT-22 cells. F. In this study, a low expression model of NEDD4L was constructed by siRNA technology and then treated with CHX for 0, 4, 8, and 12 h, respectively, to detect the expression of TFRC and NEDD4L proteins to evaluate the protein half-life. G. TFRC protein ubiquitination was detected after MG132 (10 μM) treatment