Figure 6.

SLC7A1 Might Be a Ubiquitination Substrate of SPOP

(A) Venn map of upregulated proteins and downregulated ubiquitin-modified protein for the SPOP S119N group compared with the SPOP OE group. (B) Expression of SLC7A1 (substrate of SPOP) in the control, shSPOP, SPOP OE, and SPOP S119N groups was detected by western blot analysis. (C) Real-time PCR was used for the detection of relative SLC7A1 mRNA level in the control, shSPOP, SPOP OE, and SPOP S119N groups. (D) A coimmunoprecipitation (coIP) assay was performed for SPOP and SLC7A1, SLC7A1, and ubiquitin. (E) A co-localization assay was performed for SPOP and SLC7A1 in HepG2 and Huh6 cells (original magnification ×63). (F) Western blot analysis showed the expression of SLC7A1 with 50 μM MG132 treatment after 0, 0.75, 1.5, 3, and 6 h. (G) Western blot analysis showed the expression of SLC7A1 with 0.1 μM cycloheximide after 0, 0.75, 1.5, 3, 6, and 12 h in the control, SPOP OE, and SPOP S119N groups, respectively.