Fig. 4: USP32 directly interacts with PKM2. | Cell Death & Disease

Fig. 4: USP32 directly interacts with PKM2.

From: USP32 promotes temporomandibular joint osteoarthritis by modulating PKM2 stability and glycolytic metabolism in chondrocytes

Fig. 4

A Relative sequence coverage of top metabolic proteins interacting with USP32 detected by immunoprecipitation combined with mass spectrometry. B Immunoprecipitation validation of the interaction between USP32 and the top proteins identified in (A). C Mass spectrometric analysis of immunoprecipitated proteins. D Co-immunoprecipitation of USP32 and PKM2 in MCCs under inflammatory and normal conditions using anti-USP32 antibody and immunoglobulin G (IgG). E Co-immunoprecipitation of USP32 and PKM2 in MCCs under inflammatory and normal conditions using anti-PKM2 antibody and IgG. F Immunofluorescence staining of USP32 (red) and PKM2 (green) in MCCs; the quantification of fluorescence intensity was based on the white line in the control group (G) and IL-1β group (H). Scale bar 10 μm. I Coimmunoprecipitation of USP32 and PKM2 in HEK-293T cells co-transfected with Flag-USP32 and Myc-PKM2 overexpression plasmids. USP32 was immunoprecipitated by anti-Flag antibody. J PKM2 was immunoprecipitated by anti-Myc antibody. K AlphaFold prediction of the molecular docking between USP32 and PKM2. L The schematic of the PKM2 truncated mutation is shown on the left. M Immunoprecipitation of Flag-USP32 and Myc-tagged PKM2 truncated mutant or full-length plasmids co-transfected into HEK-293T cells using anti-Myc antibody.

Back to article page