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. 2020 Jan 23;11:438. doi: 10.1038/s41467-020-14332-x

Fig. 4. PPARγ directly promotes transcription of Srebf1 in cooperation with PLZF.

Fig. 4

a Influences of T007 on mature and immature forms of SREBP1 and SREBP2 in expanded iNKT cells after activating overnight. b Total SREBP1 protein in iNKT cells measured by flow cytometry. c–e Effects of Pparg shRNA or Srebf1 shRNA on SREBP1 expression (c), Fillipin III staining (d), and percentages of IFN-γ+ iNKT cells after activation (e). f, g SREBP1 expression (f) and Fillipin III staining (g) in iNKT from PLZF-cre Ppargfl/fl mice and Ppargfl/fl mice. h, i Fragments in Srebf1 promoter region (h) and binding of these fragments to PPARγ measured by ChIP-qPCR (i). j Co-immunoprecipitation of PPARγ and PLZF in expanded iNKT cells with indicated treatments. (k, l) Co-localization (k) and co-localization coefficient (l) of PPARγ and PLZF in iNKT cells with indicated treatments. Bar, 5 μm. m mRNA of Pparg1 and Pparg2 in iNKT, CD4+ T, and CD8+ T cells. n Expression of PPARγ, PLZF, and SREBP1 in 293 T cells overexpressing PPARγ1, PPARγ2, PLZF, PPARγ vector (γ.V.), and PLZF vector (PLZF.V.) alone or in combination. o, p Total SREBP1 protein measured by flow cytometry (o) and mRNA of Srebf1 (p) in 293 T cells described in n. (q) Total SREBP1 protein in iNKT cells co-overexpressing PPARγ1 and PLZF, or co-overexpressing PPARγ2 and PLZF. r Co-immunoprecipitation of PPARγ and PLZF in 293 T cells co-overexpressing PPARγ1 and PLZF, or co-overexpressing PPARγ2 and PLZF. s Binding of PPARγ to putative PPRE (ATCTCACAGGTCA) at Srebf1 promoter region in 293T cells overexpressing PPARγ2 and PLZF as indicated. Data are representative of three independent experiments (a, j, k, n, r), or are means ± SEM of three (c, d) or four independent experiments (m, o, q), nine (b) or six biological replicates (i, p, s), or six mice (f, g), pooled from three independent experiments. Data were analyzed by unpaired Student’s t-test (b, l) or Mann–Whitney test (c–g, i, m, o, p, q, s). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.