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. 2021 Jul 26;12(8):737. doi: 10.1038/s41419-021-04028-5

Fig. 6. WTAP phosphorylation at serine 341 by ERK1/2 stabilizes WTAP to promote breast cancer cell glycolysis.

Fig. 6

A Co-IP of WTAP with p-ERK1/2 (Thr202/Tyr204) in whole-cell extracts from MCF-7C5RN cells with or without ERK1/2 inhibition (left). Quantitative analysis of co-interaction between WTAP and p-ERK1/2 (right). B Co-IP of p-ERK1/2 (Thr202/Tyr204) with WT or S341A mutated Flag-WTAP in whole-cell extracts from MCF-7C5RN cells treated with or without 10 μM CGP57380. The phosphoserine (p-Serine) of immunoprecipitated Flag-WTAP were detected using pan-phosphoserine antibody. C Immunoblotting of phosphoserine (p-Serine) of immunoprecipitated WTAP in MCF-7C5RN and MDA-MB-231C5RN cells with or without ERK1/2 inhibition. D MCF-7C5RN cells were cultured by CHX (10 mg/mL) for 0–12 h. Lysates were used to measure the protein levels of FLAG. The relative fold is indicated and plotted in the right panel. E Lactate production was examined in MCF-7 and MDA-MB-231 cells transfected with vector, wild-type, or mutant WTAP (S341A). F Analysis of ECAR in MCF-7 and MDA-MB-231 cells transfected with vector, wild-type, or mutant WTAP (S341A). G Effects of breast cancer cells with C5RN culture and shWTAP transfection on tumor growth using mice model. The volumes are measured every 7 days. Data represent the mean ± SD of at least three independent experiments. ***P < 0.001, Ctr-N C5aR1-negative neutrophils.