Fig. 6

SULT2B1 regulates the transcription of PKM2 by c-MYC and promotes the degradation of PKM2 by autophagy. A The expression of c-MYC was monitored in the indicated cells by immunoblotting. B The JASPAR database was used to predict the binding sequences of c-MYC in the promoter of PKM2. C The ChIP assay was performed, and the DNA bound to c-MYC was subjected to PCR using the specific primer, followed by agarose gel electrophoresis analysis. D Total ubiquitin was detected in both KO-Ctrl and KO-SULT2B1 HT29 cells by immunoblotting. E The co-immunoprecipitation was conducted with the primary antibody of PKM2, and the immunoprecipitate was subjected to SDS-PAGE electrophoresis analysis. F and G Immunoblotting and RT-PCR were used to identify the particular molecules involved in autophagy. H Using immunoblotting, the P62 and LC3 contents in KO-Ctrl and KO-SULT2B1 cells treated with or without chloroquine (CQ) were examined. I and J Indicated cells were transfected with the mCherry-GFP-LC3 adenovirus, and then exposed to indicate treatment for 2 h. The pictures were obtained by fluorescence microscope. K Immunofluorescence was performed with indicated antibodies. (L) The content of PKM2 in the indicated cells, either with or without CQ treatment, was determined by immunoblotting