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. 2022 Nov 17;30(2):383–396. doi: 10.1038/s41418-022-01090-0

Fig. 4. ARID2 promotes the ubiquitination of JAK2 via the E3 ligase NEDD4L.

Fig. 4

A Relative mRNA levels of JAK2 in the livers of WT and Arid2 LKO mice (upper panel, n = 6 per group) and in control and ARID2 knockdown HepG2 cells (lower panel). The mRNA expression levels were normalized to that of Gapdh/GAPDH. B, C Ubiquitination of JAK2 was examined in 293T, Hep3B (B) and primary hepatocytes (C). Cells were treated with 10 μM MG132 for 8 h. IgG was used as a negative control. D Network view of the E3 ligases for JAK2 predicted by Ubi browser website. E Relative mRNA (n = 6 per group) and protein levels of Nedd4l in the liver tissues of WT and Arid2 LKO mice. The mRNA expression level was normalized to that of Gapdh. F Ubiquitination of JAK2 was examined in 293T cells transfected with vector, WT and mutant NEDD4L, respectively. Cells were treated with 10 μM MG132 for 8 h. IgG was used as a negative control. ΔHECT, deletion of HECT domain. G–I Co-IP was performed to examine the interaction between endogenous JAK2 and NEDD4L in Hep3B cells, primary hepatocytes and HepG2 cells. IgG was used as a negative control. All experiments were repeated 3 times independently. All data were analyzed by two-tailed unpaired Student’s t test and represented as the mean ± SEM. *p < 0.05, ns not significant.