Journal: Cell proliferation

Article Title: Scutellarin Alleviates Bone Marrow Mesenchymal Stromal Cellular Senescence via the Ezh2-Nrf2 Signalling Axis in Diabetes-Induced Bone Loss.

doi: 10.1111/cpr.13790

Figure Lengend Snippet: FIGURE 3 | SCU suppresses diabetes-induced senescence of LepR+ MSCs by upregulating Nrf2. p16tdTom DOP mice were treated with citrate buf- fer or SCU daily for 8 weeks. (A) Immunofluorescence staining of femur sections using antibodies against LepR. Red: tdTom+ cells; Green: LepR+; DAPI stains nuclei blue. (B) Quantitative analysis of the percentage of LepR+tdTom+ cells among all LepR+ primary trabecular cells per mm2 tissue area (% tdTom+ LepR+ cells/Ar). (C) Representative images of the flow cytometry analysis. (D) The relative fold changes in the number of tdTom and LepR double-positive cells in the vehicle and SCU groups. (E) Representative immunoblotting images showing the protein expression levels of Nrf2. (F) Representative images of EdU staining. (H) Representative images of in vitro SA-βGal staining of CD45−tdTom+ cells. (G and I) Quantification of the percentage of cells that expressed EdU and SA-βGal relative to the total number of sorted LepR+ CD45- cells. (J–N) Representative qRT–PCR analyses of p21, p16, Ki67, TNF-α and IL-6 mRNA expression in CD45−LepR+ cells. Ar, tissue area; tdTom, tdTomato. *p < 0.05, **p < 0.01 by one- way ANOVA.

Article Snippet: Nrf2+/− mice, LepR- Cre mice, Ezh2flox/flox mice, and Rosa- LSL- tdTomato mice were obtained from Cyagen Biosciences (Guangzhou, China).

Techniques: Immunofluorescence, Staining, Flow Cytometry, Western Blot, Expressing, In Vitro, Quantitative RT-PCR

Journal: Cell proliferation

Article Title: Scutellarin Alleviates Bone Marrow Mesenchymal Stromal Cellular Senescence via the Ezh2-Nrf2 Signalling Axis in Diabetes-Induced Bone Loss.

doi: 10.1111/cpr.13790

Figure Lengend Snippet: FIGURE 5 | SCU upregulated Nrf2 via the repression of Keap1 by recruiting Ezh2. (A) Representative immunoblotting images showing the pro- tein expression levels of Nrf2. (B-E) Representative qRT–PCR analyses of HO-1, NQO-1, Nrf2, and Keap1 mRNA expression in CD45−LepR+ cells. (F) Representative immunoblotting images showing the protein expression levels of Keap1. (G) ChIP and input DNA were measured using real-time PCR with specific primers targeting the promoter regions of the indicated genes. (H) Representative qRT–PCR analyses of Ezh2 mRNA. (I) Relative Keap1 mRNA levels were compared in SCU-treated LepR+MSCs transfected with EZH2 siRNA or control siRNA. (J) Representative immunoblot- ting images showing the protein expression levels of Ezh2 and H3K27me3. (K,M) We crossed the LepR-Cre strain with the Rosa-LSL-tdTom strain. Representative confocal images of immunofluorescence staining of femur sections. Red: TdTom+ cells; Green: Ezh2+ or H3K27me3+ cells; DAPI stains nuclei blue. (L,N) Quantification of the percentage of tdTom-expressing Ezh2+ cells and H3K27me3+ cells to all tdTom+ cells. tdTom, tdto- mato. *p < 0.05, **p < 0.01.

Article Snippet: Nrf2+/− mice, LepR- Cre mice, Ezh2flox/flox mice, and Rosa- LSL- tdTomato mice were obtained from Cyagen Biosciences (Guangzhou, China).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Transfection, Control, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Leptin signaling in the dorsomedial hypothalamus couples breathing and metabolism in obesity

doi: 10.1016/j.celrep.2023.113512

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: In a separate group LepR b - Cre DIO mice of animals, we administered a retrograde Cre -dependent caspase virus AAV2-retro.EF1a.Flex.taCasp3.T2A.TEVp.WPRE.hGH (7×10 12 vg.mL −1 Penn Vector Core – Addgene, #45580) in the dorsal raphe (DR) using the following stereotactic coordinates from the animal’s bregma: −4.6mm caudal and −3.0 ventral.

Techniques: Virus, Plasmid Preparation, Recombinant, Software