PPARγ Mediates Transdifferentiation of CX3CR1+-Derived Cells into Adipocytes

Figures

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  Figure 1. RFP+SVF cells in adipose tissue of Cx3cr1cre: Rosa26Td mice. (A) Representative images of RFP+ cells in BAT and sWAT. (B) Quantification of RFP+ cell ratio in BAT and sWAT. n = 6 per group. (C) Representative images of SVF cells from BAT and sWAT. (D) Quantification of RFP+ cell area ratio of SVF cells in BAT and sWAT. n = 6 per group. (E) Quantification of SVF cells’ size in BAT and sWAT. n = 10 cells per group. Scale bars: 50 μm. Values represent mean ± SEM. Student’s t-tests (B,D,E), *** p < 0.001, ** p < 0.01, * p < 0.05.

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  Figure 2. RFP+SVF cells transdifferentiate into adipocytes in BAT and sWAT of Cx3cr1cre: Rosa26Td mice. (A) Representative images of RFP+SVF cells differentiated into adipocytes in BAT of Cx3cr1cre: Rosa26Td mice. (B) Quantification of RFP+ area ratio in BAT of Cx3cr1cre: Rosa26Td mice. (C) Representative images of RFP+SVF cells differentiated into adipocytes in sWAT of Cx3cr1cre: Rosa26Td mice. (D) Quantification of RFP+ area ratio in sWAT of Cx3cr1cre: Rosa26Td mice. n = 6 per group. Scale bars: 2 μm. Values represent mean ± SEM. Student’s t-tests (B,D), * p < 0.05, ns = no significant.

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  Figure 3. Cold exposure promotes lipogenesis of adipocytes in BAT, sWAT, eWAT, and iWAT of Cx3cr1cre: Rosa26Td mice. (A) H&E staining of BAT, sWAT, eWAT, and iWAT under cold exposure. (B) Quantification of adipocyte size of BAT, sWAT, eWAT, and iWAT under cold exposure. n = 8 cells per group. Scale bars: 100 μm. Values represent mean ± SEM. Student’s t-tests (B), **** p < 0.0001, *** p < 0.001.

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  Figure 4. Cold exposure promotes lipogenesis of RFP+ cells in BAT, sWAT, eWAT, and iWAT of Cx3cr1cre: Rosa26Td mice. (A) Representative images of Plin1 expression of RFP+ cells in BAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (B) Quantification of RFP+ Plin1+ area in BAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (C) Representative images of Plin1 expression of RFP+ cells in sWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (D) Quantification of RFP+ Plin1+ area in sWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (E) Representative images of Plin1 expression of RFP+ cells in eWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (F) Quantification of RFP+ Plin1+ area in eWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (G) Representative images of Plin1 expression of RFP+ cells in iWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. (H) Quantification of RFP+ Plin1+ area in iWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. n = 8 per group. Scale bars: 100 μm. Values represent mean ± SEM. Student’s t-tests (B,D,F,H), *** p < 0.001, ** p < 0.01, * p < 0.05.

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  Figure 5. Cold exposure promotes CX3CR1 and PDGFRα expression of RFP+ cells in BAT and sWAT of Cx3cr1cre: Rosa26Td mice. (A) Flow cytometric analysis of expression of CX3CR1 and PDGFRα by RFP+ SVF cells in BAT and sWAT of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. Quantification of CX3CR1+ and PDGFRα+ ratio by RFP+ SVF cells in BAT (B) and sWAT (C) of Cx3cr1cre: Rosa26Td mice under RT and cold exposure. n = 5 per group. Values represent mean ± SEM. Student’s t-tests (B,C), ** p < 0.01, * p < 0.05.

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  Figure 6. HFD inhibits lipogenesis of RFP+ cells in BAT, sWAT, and eWAT of Cx3cr1cre: Rosa26Td mice. (A) Representative images of Plin1 expression in BAT of Cx3cr1cre: Rosa26Td mice under HFD. (B) Quantification of RFP+ Plin1+ area in BAT of Cx3cr1cre: Rosa26Td mice under HFD. (C) Representative images of Plin1 expression in sWAT of Cx3cr1cre: Rosa26Td mice under HFD. (D) Quantification of RFP+ Plin1+ area in BAT of Cx3cr1cre: Rosa26Td mice under HFD. (E) Representative images of Plin1 expression in eWAT of Cx3cr1cre: Rosa26Td mice under HFD. (F) Quantification of RFP+ Plin1+ area in BAT of Cx3cr1cre: Rosa26Td mice under HFD. n = 7 per group. Scale bars: 100 μm. Values represent mean ± SEM. Student’s t-tests (B,D,F), ** p < 0.01. * p < 0.05.

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  Figure 7. GW9662 inhibits the lipogenic differentiation of RFP+SVF cells in BAT and sWAT of Cx3cr1cre: Rosa26Td mice. (A) Representative images of Plin1 expression of RFP+ SVF cells differentiated into brown adipocyte (BAT-B) and white adipocyte (BAT-W) after DMSO or GW9662 treatment. (B) Quantification of RFP+ Plin1+ area of BAT-B and BAT-W after DMSO or GW9662 treatment. (C) Quantification of Plin1+ area of BAT-B and BAT-W after DMSO or GW9662 treatment. (D) Representative images of Plin1 expression of RFP+ SVF cells differentiated into brown adipocyte (sWAT-B) and white adipocyte (sWAT-W) after DMSO or GW9662 treatment. (E) Quantification of RFP+ Plin1+ area of sWAT-B and sWAT-W after DMSO or GW9662 treatment. (F) Quantification of Plin1+ area of sWAT-B and sWAT-W after DMSO or GW9662 treatment. n = 6 per group. Scale bars: 50 μm. Values represent mean ± SEM. Two-way ANOVA (B,C,E,F), *** p < 0.001, ns = no significant.