Fig 1: Secretome profile of HBPCs. A Timeline of CM collection. HBPCs were treated with LPS and/or PDGF-BB for 1 h, washed, cultured for 24 h, and CMs were collected. B Heat map of different protein levels in HBPC CM. The secreted levels of each factor are expressed as fold changes from the amount secreted in PBS CM. Colors from blue to red indicate a low to high protein level. C The top 12 proteins secreted in response to the stimulation are summarized in the graph. D Effects of inhibitors on the inflammatory priming of BMDMs by the pericyte secretome. BMDMs were pre-incubated with 1 μM SB225002 (CXCR1/2 inhibitor), 5 μM RS102895 (CCR2 inhibitor), or 100 nM C021 (CCR4 inhibitor) in serum-free RPMI 1640 media for 1 h, followed by a treatment with PBS CM or LPS + PDGF-BB CM for 12 h. Cells were then stimulated with 10 ng/mL LPS for 24 h, and Nos2 expression was measured as an index of inflammatory priming (n = 5). E mRNA level of CXCL5 in HBPCs treated with 100 ng/mL LPS and 100 ng/mL recombinant human PDGF-BB for 3 h (n = 6). F mRNA level of CXCL5 in HBPCs treated with 100 μM Palmitate (Pal) and 100 ng/mL recombinant human PDGF-BB for 6 h (n = 6). G Cxcl5 mRNA expression in the hypothalamus of FL and Pdgfrb∆SYS-KO fed HFD for 4 weeks (n = 12–13). Data are presented as the mean ± SEM. *p < 0.05 and **p < 0.01, significantly different between each group
Fig 2: PDGF signaling mediates chronic inflammation in the hypothalamus. A Representative confocal images of cFos (green) and POMC (red) immunoreactivities and counterstaining with Hoechst 33342 (blue) in the ARC of FL and Pdgfrb∆SYS-KO fed HFD for 4 weeks (n = 7–8). Scale bar = 200 μm. The right two panels are high magnification of the gray line square area. White arrows indicate Hoechst+cFos+POMC+ cells. Scale bar = 50 or 20 μm. The ratio of POMC-positive neurons in Hoechst-positive cells and cFos-POMC double-positive neurons in POMC-positive neurons are quantified. B Representative confocal images of UCP1 immunoreactivity (green) and Hoechst 33,342 (blue) staining in the BAT of FL and Pdgfrb∆SYS-KO fed HFD for 4 weeks. Scale bar = 100 μm. Mean fluorescent intensity (MFI) of UCP1 in the BAT (n = 8). C mRNA levels in the hypothalamus of C57BL6J mice fed a normal chow diet or HFD for 4 weeks (n = 8). D mRNA levels in the hypothalamus of FL and Pdgfrb∆SYS-KO fed HFD for 4 weeks (n = 12–13). E Representative confocal images of Iba1 immunoreactivity (green) in the ARC and VMH of FL and Pdgfrb∆SYS-KO fed HFD for 4 weeks (n = 8–12). Scale bar = 100 μm. Each region was indicated by a dotted line. The numbers of Iba1-positive microglia in the ARC and VMH were quantified. Data are presented as the mean ± SEM. *p < 0.05 and **p < 0.01
Fig 3: PDGF signaling in pericytes, but not in neurons, mediates the dysfunction of energy metabolism. A Pdgfb and Pdgfrb mRNA levels in the hypothalamus of Pdgfrb∆SYS-KO fed HFD for 4 weeks (n = 12–13). B Body weights of Pdgfrb∆SYS-KO (n = 12–13). C–E Metabolic parameters of Pdgfrb∆SYS-KO fed HFD for 3–4 weeks. Energy expenditure (C), locomotor activity (D) (n = 9–12), and food intake (E) (n = 4–5). F Pdgfrb mRNA levels in the hypothalamus of Pdgfrb∆CaMKII-KO fed HFD for 4 weeks (n = 8–10). G Body weights of Pdgfrb∆CaMKII-KO (n = 8–10). H Energy expenditure in Pdgfrb∆CaMKII-KO fed HFD for 3 weeks (n = 8–10). Data are presented as the mean ± SEM. *p < 0.05 and **p < 0.01
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