Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: Chemerin and RvE1 differentially regulated mTORC1 signaling via CMKLR1. (a) Gene set enrichment analysis (GSEA) analysis for gene signatures of mTOR signaling pathway in iWAT from Rarres2 KO compared with C57BL/6J control mice. NES, normalized enrichment score. FDR, false discovery rate. (b, c) Western-blot analysis for level of p-S6K and S6K protein in undifferentiated 3T3-L1 treated with 100 ng/ml chemerin (b) or 100 ng/ml RvE1 (c). (d, e) Western-blot analysis for level of UCP1, p-S6K and S6K protein in 3T3-L1 mature adipocytes treated with 1 µM CL, 1 µM CL with 100 ng/ml chemerin (CL+CHE), 10 µM Mi or 100 ng/ml (0.285 µM) RvE1. (f) C57BL/6J mice fed with NCD for 8 weeks were injected with Vehicle, chemerin-9 (1 µg/mouse/day), Rapamycin (2 mg/kg/day) or Rapamycin (2 mg/kg/day) with chemerin-9 (1 µg/mouse/day) over 7 days. Western-blot analysis for level of UCP1 protein in iWAT. (g) C57BL/6J mice fed with NCD for 8 weeks were injected with Vehicle, RvE1 (1 µg/mouse/2day), Rapamycin (2 mg/kg/2day) or Rapamycin (2 mg/kg/2day) with RvE1 (1 µg/mouse/2day) over 14 days. Western-blot analysis for level of UCP1 protein in iWAT. (h-i) Western-blot analysis for level of p-S6K, S6K, UCP1 and CMKLR1 protein in different 3T3-L1 cell pools treated with 100 ng/ml chemerin, 10 µM Mirabegron or 100 ng/ml RvE1 as well as transfected with Cas9 and guide RNA targeting the Cmklr1 gene sequence. gNS indicates 3T3-L1 cell pools transfected with Cas9 only and g#1 indicates 3T3-L1 cell pools transfected with Cas9 and guide RNA. The ImageJ software was used for gray scanning.

Article Snippet: The Cas9 lentivirus vector and gRNA1/2 lentivirus vector were purchased from Cyagen Biosciences and transduced to 3T3-L1 cells.

Techniques: Control, Western Blot, Injection, Transfection, Sequencing, Software