Fig. 2

RA sustained the intracellular level of Asc. a The protein levels of Glut1 and Glut3 were determined by immunoblotting on day 6 during reprogramming with AscPNa. b-c The bind sites of RARA::RARG were identified on the promoters of Glut1 and Glut3 by Pscan software. Promoter assay confirmed the abilities of RA to affect the functions of the promoters of Glut1 and Glut3 in MEFs. d The intracellular level of Asc was determined on day 6 during reprogramming with N2B27 medium (24 h after replacing medium with fresh medium containing AscPNa). e-j MEFs were cultured with N2B27 medium containing 0.16 mM AscPNa or equivalent concentration of DHAA (E&H). Different concentrations of RA were also used. The intracellular levels of Asc was determined at different time points during the treatment with AscPNa (F&I) or DHAA (f), and the area under curve (intracellular levels of Asc multiple time length) were summarized in (g) and (j) respectively. k-n Different concentrations of RA were used to treat MEFs cultured in N2B27 medium in the presence or absence of 0.16 mM AscPNa (k). The expression of Glut1 (l), Glut3 (m), and Svct2 (n) was determined by qPCR. The comparisons were preformed between RA-treated groups and corresponding control (DMSO) groups or between indicated groups with one-way ANOVA except with two-way ANOVA in (c, f & j). Experiments were repeated for at least five times (n ≥ 5). Standard deviations were provided