Journal: Clinical and Translational Medicine

Article Title: CRISPRa‐based activation of Fgf21 and Fndc5 ameliorates obesity by promoting adipocytes browning

doi: 10.1002/ctm2.1326

Figure Lengend Snippet: Conditioned media of muscle cells with myokines activation promote adipocytes browning in vitro. (A) Western blot assay shows the level of UCP1 protein in 3T3‐L1 preadipocytes and 3T3‐L1 derived adipocytes with activation of Fgf21 / Fndc5 / Fgf21 + Fndc5 . Tubulin was used as an internal reference. (B–D) qRT‐PCR quantification of BAT‐selective genes ( Cidea , Elovl3 ), mitochondrial genes ( cyc , cox8b , cox7a1 ), adipocyte markers ( Pparg , Tfap2a , Adipoq ) and thermogenesis‐related genes ( Ucp1 , Ppargc1a ) in 3T3‐L1 preadipocytes and 3T3‐L1‐derived adipocytes treated with conditioned media of Fgf21 / Fndc5 / Fgf21 + Fndc5 ‐activated C2C12 cells.

Article Snippet: The primary antibodies involving against FGF21 (1:1000; Abcam; Cat# ab171941, RRID: AB_2629460), FNDC5 (1:1000; Proteintech; Cat# 23995‐1‐AP, RRID: AB_2879394), UCP1 (1:1000; ABclonal; Cat# A5857, RRID: AB_2766607), β‐actin (1:5000; ABclonal; Cat# AC038, RRID: AB_2863784), α‐tubulin (1:5000; ABclonal; Cat# AC007, RRID: AB_2772755) and HRP conjugated goat anti‐mouse and goat anti‐rabbit secondary antibody (1:10 000; Jackson ImmunoResearch) were used.

Techniques: Activation Assay, In Vitro, Western Blot, Derivative Assay, Quantitative RT-PCR

Journal: Clinical and Translational Medicine

Article Title: CRISPRa‐based activation of Fgf21 and Fndc5 ameliorates obesity by promoting adipocytes browning

doi: 10.1002/ctm2.1326

Figure Lengend Snippet: In vivo CRISPRa‐based activation of myokines promotes adipocytes browning in DIO mice. (A) H&E staining shows reduced lipid content in BAT of mice with CRISPRa‐based myokines activation at 4‐ and 8‐weeks post‐injection. Images are representative of four sections of three mice in each group. (B) Immunohistochemical staining for UCP1 (BAT marker) of BAT sections from chow or HFD mice that received AAV9‐CRISPRa vectors. (C) Western blot analysis of the UCP1 protein content in BAT from mice in each group. (D) The histogram depicts the densitometric analysis of immunoblots in C. (E) qRT‐PCR quantification of markers specific to BAT ( Elovl3 , Cidea ) and thermogenesis ( Ucp1 , Ppargc1a ) in BAT of mice in each group. The mRNA levels were normalised to sgSCR group. Scale bar in A and B, 100 μm.

Article Snippet: The primary antibodies involving against FGF21 (1:1000; Abcam; Cat# ab171941, RRID: AB_2629460), FNDC5 (1:1000; Proteintech; Cat# 23995‐1‐AP, RRID: AB_2879394), UCP1 (1:1000; ABclonal; Cat# A5857, RRID: AB_2766607), β‐actin (1:5000; ABclonal; Cat# AC038, RRID: AB_2863784), α‐tubulin (1:5000; ABclonal; Cat# AC007, RRID: AB_2772755) and HRP conjugated goat anti‐mouse and goat anti‐rabbit secondary antibody (1:10 000; Jackson ImmunoResearch) were used.

Techniques: In Vivo, Activation Assay, Staining, Injection, Immunohistochemical staining, Marker, Western Blot, Quantitative RT-PCR

Journal: Pharmaceuticals

Article Title: Rutaecarpine Promotes Adipose Thermogenesis and Protects against HFD-Induced Obesity via AMPK/PGC-1α Pathway

doi: 10.3390/ph15040469

Figure Lengend Snippet: Rut increases expression of PGC-1α and drives the thermogenesis program. ( A ) The screening result of Rut in activating PGC-1α. ( B ) Effect of Rut on activating PGC-1α in C3H10-T1/2. ( C ) RT-qPCR analysis of genes indicated in C3H10-T1/2. ( D ) Expressions of protein indicated in C3H10-T1/2 treated by Rut for 24 h. ( E – G ) Relative protein level of PGC-1α ( E ), UCP1 ( F ) and NRF2 ( G ) compared to β-actin. n = 3 per group. Data are presented as the means ±SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, Rut groups versus vehicle group by one-way ANOVA.

Article Snippet: The antibody sources were as follows: UCP1 (Abclonal, A5857), PGC1α (Calbiochem, ST1202); AMPKα (Cell Signaling Technology, #2532); phospho-AMPKα (Thr172) (Cell Signaling Technology, #2535); ACC (Cell Signaling Technology, #3662); phospho-ACC (Ser79) (Cell Signaling Technology, #3661); and β-actin (Abgent, AM1021B).

Techniques: Expressing, Quantitative RT-PCR

Journal: Pharmaceuticals

Article Title: Rutaecarpine Promotes Adipose Thermogenesis and Protects against HFD-Induced Obesity via AMPK/PGC-1α Pathway

doi: 10.3390/ph15040469

Figure Lengend Snippet: Rut stimulates thermogenic program via AMPK/PGC-1α pathway. ( A ) Expression of PGC-1α and UCP1 protein by siRNA against PGC-1α. ( B , C ) Relative protein level of PGC-1α ( B ) and UCP1 ( C ) compared to β-actin. ( D ) The mRNA level of Pgc-1α and Ucp1 after blockage of PGC-1α. ( E ) Expression of p-AMPK, AMPK, p-ACC and ACC protein in C3H10-T1/2 treated by Rut. ( F ) Expression of PGC-1α and UCP1 protein in C3H10-T1/2 with siRNA against AMPK and Rut treatment. ( G , H ) Relative protein level of PGC-1α ( G ) and UCP1 ( H ) compared to β-actin. ( I ) The mRNA level of Pgc-1α and Ucp1 after silence of AMPK. n = 3 per group. Data are presented as the means ±SEM. * p < 0.05, ** p < 0.01, Rut group versus vehicle group; # p < 0.05, ## p < 0.01, ### p < 0.001, siRNA group versus control group by Student’s t -test.

Article Snippet: The antibody sources were as follows: UCP1 (Abclonal, A5857), PGC1α (Calbiochem, ST1202); AMPKα (Cell Signaling Technology, #2532); phospho-AMPKα (Thr172) (Cell Signaling Technology, #2535); ACC (Cell Signaling Technology, #3662); phospho-ACC (Ser79) (Cell Signaling Technology, #3661); and β-actin (Abgent, AM1021B).

Techniques: Expressing