Fig. 3: Butyrate-induced SLC7A11 downregulation via the GSK3β-β-TRCP1-NRF2 pathway.

HT29 cells were treated with the indicated concentrations of NaB or in combination with bafilomycin A1 for 6 h, the levels of the indicated proteins were evaluated by WB (A), the expression of NRF2 (B) and SLC7A11 (C) were measured using qRT-PCR. HT29 cells were treated with CHX for the indicated time after pretreatment with NaB for 6 h, and then WB was used to evaluate the levels of NRF2 (D). Quantitative data for NRF2 protein levels are presented (E). F–H β-TRCP1-knockdown HT29 cells were treated with the indicated concentrations of NaB for 6 h, and then WB was used to evaluate the levels of NRF2 (F) and quantitative data for NRF2 protein levels are presented (H). The knockdown efficiency was measured by qRT-PCR (G). I β-TRCP1-knockdown HT29 cells were treated with CHX for the indicated time after pretreatment with NaB for 6 h, and then WB was used to evaluate the levels of NRF2. The knockdown efficiency was shown in (G). J β-TRCP1-knockdown HT29 cells were treated with the indicated concentrations of NaB for 6 h, and the expression of SLC7A11 was measured using qRT-PCR. The knockdown efficiency was shown in (G). K, L HT29 cells were treated with CHX for the indicated time after pretreatment with NaB or in combination with LiCl (10 mM) for 6 h, and then WB was used to evaluate the levels of NRF2 (K). Quantitative data for NRF2 protein levels are presented (L). M, N HT29 cells were treated with the indicated concentrations of NaB or in combination with LiCl, and then WB was used to evaluate the levels of NRF2 (M). Quantitative data for NRF2 protein levels are presented (N). O–Q GSK3β-knockdown HT29 cells were treated with the indicated concentrations of NaB for 6 h, and then WB was used to evaluate the levels of NRF2 (O). Quantitative data for NRF2 protein levels are presented (Q) and the knockdown efficiency was measured by qRT-PCR (P). GSK3β-knockdown HT29 cells were treated with CHX for the indicated time after pretreatment with NaB for 6 h, and then WB was used to evaluate the levels of NRF2 (R), quantitative data for NRF2 protein levels are presented (S). The knockdown efficiency was shown in (P). T GSK3β-knockdown HT29 cells were treated with the indicated concentrations of NaB for 6 h, and the expression of SLC7A11 was measured using qRT-PCR. The knockdown efficiency was shown in (P).