Fig. 3: The mechanism by which WTAP overexpression enhanced forkhead box other 1 (Foxo1) expression. | Laboratory Investigation

Fig. 3: The mechanism by which WTAP overexpression enhanced forkhead box other 1 (Foxo1) expression.

From: The N6-methyladenosine writer WTAP contributes to the induction of immune tolerance post kidney transplantation by targeting regulatory T cells

Fig. 3

A–E all performed on CD4+ T cells generated from the whole-blood samples of IR. A RNA Immunoprecipitation (RIP) and methylated RIP (Me-RIP) assays were performed on cells. The enrichment of Foxo1 mRNA in the products immunoprecipitated by the anti-WTAP or anti-m6A antibody was examined using qRT-PCR. IgG was used as a negative control. B Me-RIP assay was performed on cells infected with Lenti-WTAP or Lenti-control. The enrichment of Foxo1 mRNA in the products immunoprecipitated by the anti-m6A antibody was examined using qRT-PCR. C mRNA and protein levels of Foxo1 in cells infected with Lenti-WTAP or Lenti-control were measured using qRT-PCR and Western blot, respectively. β-actin was used as an internal control. D Cells were infected with Lenti-WTAP or Lenti-control and then treated with cycloheximide (CHX, 10 μM) and the Foxo1 protein level was examined using Western blot at indicated time points after CHX treatment. E Cells were infected with Lenti-WTAP or Lenti-control, and polysome fractionation assay was conducted to measure relative distribution of Foxo1 mRNA. *P < 0.05, **P < 0.01 vs. IgG or Lenti-control. ns not statistically significant.

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