Figure 6.

RNF126 regulates the protein-expression levels of PDK1, PDK3 and PDK4. (a) Schematic illustration of pyruvate conversion to lactate or acetyl-CoA. (b) Western blot analysis of PDKs and LDHA expression in control and RNF126-depleted MDA-MB-231 cells cultured in the attached or detached state. Results of the densitometric analysis of bands in western blots are presented. (c) Western blot analysis of PDH-E1α and its phosphorylation levels (pSer²³², pSer²⁹³ and pSer³⁰⁰) in control and RNF126-depleted cells cultured in the attached or detached state. (d) The PDK inhibitor DCA restored ATP levels in RNF126-depleted cells. Error bars indicate the s.d. (n=3). The data were analyzed using a t-test. *P<0.05; **P<0.01. (e) Exposure to the proteasomal inhibitor MG132 eliminated differences in the levels of the PDK1, PDK3 and PDK4 proteins between control and RNF126-KD cells. The data shown in (b–e) are representative of three independent experiments with similar results.