Fig. 5

PFK participates in the effects of PPARγ agonists through associating FAO with UDP-GlcNAc/glycosylation axis. a–d The naïve CD4⁺ T cells were transfected with siCD36 (50, 100, 150, 200 nM) or siCPT1 (50, 100, 150, 200 nM), and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL) for 48 h. The mRNA levels of GFPT and PFK were measured by Q-PCR (a, b), and the activity of PFK and GFPT was determined (c, d). e The naïve CD4⁺ T cells were prepared and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (10, 30 μM), 15d-PGJ2 (3, 10 μM) as well as morin (10, 30 μM) for 48 h. The activity of PFK and GFPT was determined. f The naïve CD4⁺ T cells were transfected with siPFK, and the mRNA level of PFK was measured by Q-PCR. g–i The naïve CD4⁺ T cells were transfected with siPFK, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The intracellular level of UDP-GlcNAc was determined (g), and the level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA (h, i). j–l The naïve CD4⁺ T cells were transfected with siCD36 or siCPT1, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The activity of PFK (j) and the intracellular level of UDP-GlcNAc (k) were determined. The level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA (l). Data were presented as the means ± S.E.M. of three independent experiments (n = 3). ^##P < 0.01 versus control group; *P < 0.05, **P < 0.01 versus TGF-β group (model group); ^$P < 0.05, ^$$P < 0.01 versus rosiglitazone group; ^&P < 0.05, ^&&P < 0.01 versus 15d-PGJ2 group; ^†P < 0.05, ^††P < 0.01 versus morin group