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. 2022 Feb 24;13(2):181. doi: 10.1038/s41419-022-04622-1

Fig. 5. Ubc9 deficiency impairs PDPK1 signaling coupled with altered glycolysis.

Fig. 5

A, B Expression levels of UBC9, (p-)PDPK1, (p-)AKT, and (p-)mTOR as detected by Western blot. C OCR of control and KO CD4 T cells at basal levels, followed by sequential treatment (dashed lines) of oligomycin (Oligo), FCCP, and rotenone plus antimycin (R/A) (representative of two experiments). D Accordingly, baseline OCR (WT: 122.0 ± 14.87 pmol/min vs. KO: 70.34 ± 4.69 pmol/min, p < 0.05) (WT: n = 5, KO: n = 3), maximal respiration OCR (WT: 305.7 ± 19.29pmol/min vs. KO: 140.3 ± 33.70 pmol/min, p < 0.01) (WT: n = 5, KO: n = 5), and the reserved OCR capacity (WT: 183.7 ± 19.05 pmol/min vs. KO: 32.60 ± 12.82 pmol/min, p < 0.01) (WT: n = 5, KO: n = 3) were shown. E ECAR of control and KO CD4 T cells at basal levels, followed by sequential treatment (dashed lines) of glucose (Glc), Oligo and 2-DG (representative of two experiments). F Accordingly, baseline glycolysis (WT: 39.65 ± 2.68 mpH/min vs. KO: 17.13 ± 2.61 mpH/min, p < 0.001) (n = 5), maximal glycolytic capacity (WT: 113.2 ± 4.99 mpH/min vs. KO: 32.15 ± 7.89 mpH/min, p < 0.001) (n = 5), and glycolytic reserve (WT: 75.42 ± 3.56 mpH/min vs. KO: 12.54 ± 4.94 mpH/min, p < 0.001) were shown (n = 3). G Glucose-uptake assay was performed to measure the ability of glucose usage by WT and KO CD4 T cells (WT: 33.87 ± 1.68% vs. KO: 7.39 ± 0.44%, p < 0.001) (n = 3). H Quantification of the relative mRNA abundance of key glycolytic genes (Eno1, Glut1, Pgk1, and Pkm2) was determined by RT-PCR (n = 3, representative of two experiments). The p-value was determined by Student’s unpaired t-test.