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. 2022 Apr 8;8:176. doi: 10.1038/s41420-022-00987-6

Fig. 1. Glucose deficiency induces MAT2A-PDCD6 interaction.

Fig. 1

A Protein levels of MAT2A were detected by immunoblotting in normal cervical epithelial cells and cervical cancer cell lines. B The mass spectrometry analysis was performed in Flag tagged-MAT2A cell. MAT2A-associated proteins were listed. C MS751 cells transfected with or without 3×flag-MAT2A were cultured for 12 h under normal or glucose deprivation. Cellular extracts were subjected to immunoprecipitation with an anti-PDCD6 antibody. D C33A cells transfected with or without 3×flag-MAT2A were cultured for 12 h under normal or glucose deprivation. Cellular extracts were subjected to immunoprecipitation with an anti-PDCD6 antibody. E MS751 cells transfected with or without MAT2A shRNA were cultured for 12 h under normal or glucose deprivation. Western blotting analysis was used to determine MAT2A protein and PDCD6 protein levels (Left panel). Relative mRNA expression of PDCD6 was analyzed using RT-PCR (Right panel). F MS751 cells transfected with or without MAT2A shRNA were cultured for 12 h. The MS751 cells with MAT2A shRNA were treated with 10 μM MG132 for 6 h. Cell lysates were analyzed by Western blotting. G MS751 cells with stable expressing wild-type MAT2A or sh-MAT2A were cultured in glucose deprivation condition for 12 h, and then CHX (10 μg/ml) treatment was applied for different time courses (0, 2, 4, 6, 8 h) before harvest. The below panel showcases relative protein amounts of different groups. Error bar mean ± s.d. of triplicate experiments. ***P < 0.001, **P < 0.01, *P < 0.05. H MS751 cells with expressing Flag-MAT2A were pretreated with Compound C (10 μM), SP600125 (20 μM) and SB203580 (10 μM) for 1 h before being cultured with glucose deprivation for 12 h. Cellular extracts were subjected to immunoprecipitation with an anti-MAT2A antibody. Cell lysates were directly subjected to Western blotting.