Journal: Nutrition & Metabolism

Article Title: Differential regulation of PGC-1α expression in rat liver and skeletal muscle in response to voluntary running

doi: 10.1186/1743-7075-7-36

Figure Lengend Snippet: Total PGC-1α protein content in plantaris muscle assessed by Western blotting in sedentary control and exercised rats . Values are mean ± SE with n = 8 rats/group. * P < 0.001 vs sedentary control.

Article Snippet: The nitrocellulose membranes were washed with TBS containing 0.1% Tween-20 (TBS-T) and incubated for 4 h at 22°C with primary anti-PGC-1α antibody (polyclonal antibody of rabbit against amino acids 1-300 promoted mapped near the N-terminal portion of PGC-1 - Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Control

Journal: Nutrition & Metabolism

Article Title: Differential regulation of PGC-1α expression in rat liver and skeletal muscle in response to voluntary running

doi: 10.1186/1743-7075-7-36

Figure Lengend Snippet: Total PGC-1α protein content in soleus muscle assessed by Western blotting in sedentary control and exercised rats . Values are mean ± SE with n = 8 rats/group.

Article Snippet: The nitrocellulose membranes were washed with TBS containing 0.1% Tween-20 (TBS-T) and incubated for 4 h at 22°C with primary anti-PGC-1α antibody (polyclonal antibody of rabbit against amino acids 1-300 promoted mapped near the N-terminal portion of PGC-1 - Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Control

Journal: Nutrition & Metabolism

Article Title: Differential regulation of PGC-1α expression in rat liver and skeletal muscle in response to voluntary running

doi: 10.1186/1743-7075-7-36

Figure Lengend Snippet: Total PGC-1α protein content in liver tissue assessed by Western blotting in sedentary control and exercised rats . Values are mean ± SE with n = 8 rats/group.

Article Snippet: The nitrocellulose membranes were washed with TBS containing 0.1% Tween-20 (TBS-T) and incubated for 4 h at 22°C with primary anti-PGC-1α antibody (polyclonal antibody of rabbit against amino acids 1-300 promoted mapped near the N-terminal portion of PGC-1 - Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Control

Journal: Redox biology

Article Title: Genistein mitigates senescence of bone marrow mesenchymal stem cells via ERRα-mediated mitochondrial biogenesis and mitophagy in ovariectomized rats.

doi: 10.1016/j.redox.2023.102649

Figure Lengend Snippet: Fig. 3. Genistein promotes mitochondrial biogenesis and mitophagy in OVX-BMMSCs. (A) Protein levels of CoxIV, TOM20 and PGC1α were analyzed by western blots using β-actin as loading control (n = 3). Statistical analysis of expression of CoxIV, TOM20 and PGC1α was adjusted to ACTB. (B) The mRNA expression of Pgc1α, Tfam, Nrf1 and Tfb2m in different groups (n = 3). (C) The ATP level was measured and the values of Sham-BMMSCs were normalized to 100%. (D) Representative images of mitochondria labeled with red (Mito-Tracker Red CMXRos, MTR). The MFI of Sham-BMMSCs were normalized to 100% (n = 3). Cells were treated by genistein for 3 days. Each value is mean ± SD. Significance was calculated using a one-way ANOVA followed by the Tukey’s post-hoc test. *p < 0.05 and **p < 0.01 compared to the Sham group. #p < 0.05 and ##p < 0.01 compared to the OVX group. (E) Representative images of transmission electron microscopy (n = 3). OVX-BMMSCs were treated by genistein for 2 days. Mitochondria lost their cristae and formed large vacuoles in OVX-BMMSCs. In Sham-BMMSCs and genistein- treated OVX-BMMSCs, however, mitochondria were intact with clear cristae. Red arrow indicated the mitochondrion was engulfed in an autophagosome, and white arrowheads indicated mitochondria. Scale bar, 500 nm. (F, G) Protein levels of LC3, P62, CoxIV and TOM20 were analyzed by western blots using β-actin as loading control (n = 3). Cells were subjected to indicated treatment for 48 h. Each value is mean ± SD. Significance was calculated using a one-way ANOVA followed by the Tukey’s post-hoc test. *p < 0.05 and **p < 0.01 compared to the control group. #p < 0.05 and ##p < 0.01 compared to the genistein group. (H) Protein levels of LC3 and P62 were analyzed by western blots using β-actin as loading control (n = 3). Values represent the mean ± SD of three independent experiments. Significance was calculated using unpaired T-test. *p < 0.05 compared to the Sham group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For peroxisome proliferator-activated receptor gamma coactivator one alpha (PGC1α) and leptin receptor (LepR) immunofluorescence staining, sections were blocked with 10% goat serum (Gibco) and then incubated with primary antibodies (anti- PGC1α, 1:50, Proteintech; anti- M. Li et al. Redox Biology 61 (2023) 102649 LepR, 1:50, Invitrogen) at 4 ◦C overnight and with secondary antibodies for 1 h at 37 ◦C.

Techniques: Western Blot, Control, Expressing, Labeling, Transmission Assay, Electron Microscopy

Journal: Redox biology

Article Title: Genistein mitigates senescence of bone marrow mesenchymal stem cells via ERRα-mediated mitochondrial biogenesis and mitophagy in ovariectomized rats.

doi: 10.1016/j.redox.2023.102649

Figure Lengend Snippet: Fig. 5. Genistein attenuated senescence, promoted mitochondria biogenesis and autophagy via ERRα. OVX-BMMSCs were transfected with ERRα siRNA or control siRNA for 48 h and then cultured with 10−2 μM genistein for another 3 days. (A) Protein levels of ERRα, p53, p16INK4a and p21CIP1 and representative blots (n = 3). β-actin was used as loading control. (B) The mRNA expression of senescence related marker p53, p16INK4a and p21CIP1 in different groups (n = 3). (C, D) Repre sentative images of γH2AX staining and quantitative assessment of the γH2AX positive cells (n = 3). Scale bar, 50 μm. (E) Representative images of SA-β-gal staining and quantitative assessment of the SA-β-gal positive cells (n = 3). Scale bar, 500 μm. (F) The mRNA expression of Pgc1α, Tfam, Nrf1 and Tfb2m in different groups (n = 3). (G) The mtDNA copy number was measured in each group (n = 3). (H) Oxygen-consumption rates (OCRs) of cells at baseline and in response to oligomycin A, FCCP, and rotenone plus antimycin A. Basal respiration, ATP-linked respiration and maximal respiration were shown on the right. Data show mean ± SD of three independent experiments from 3 rats. Significance was calculated using a one-way ANOVA followed by the Tukey’s post-hoc test (A, B, D, E, F, G and H). *p < 0.05 and **p < 0.01 compared to the siNC + Control group. #p < 0.05 and ##p < 0.01 compared to the siNC + genistein group.

Article Snippet: For peroxisome proliferator-activated receptor gamma coactivator one alpha (PGC1α) and leptin receptor (LepR) immunofluorescence staining, sections were blocked with 10% goat serum (Gibco) and then incubated with primary antibodies (anti- PGC1α, 1:50, Proteintech; anti- M. Li et al. Redox Biology 61 (2023) 102649 LepR, 1:50, Invitrogen) at 4 ◦C overnight and with secondary antibodies for 1 h at 37 ◦C.

Techniques: Transfection, Control, Cell Culture, Expressing, Marker, Staining

Journal: Redox biology

Article Title: Genistein mitigates senescence of bone marrow mesenchymal stem cells via ERRα-mediated mitochondrial biogenesis and mitophagy in ovariectomized rats.

doi: 10.1016/j.redox.2023.102649

Figure Lengend Snippet: Fig. 8. Genistein treatment inhibited senescence marker p16INK4a expression and induced SIRT3 and PGC1α expression in the trabecular bone area of proximal tibia in ovariectomized rats. (A, B) Representative images of immunohistochemical staining of p16INK4a in different groups. Scale bar, 100 μM. The graph on the right showed mean intensity of p16INK4a staining (n = 9). (C, D) Representative images of immunofluorescence staining of SIRT3 (green) and nucleus (blue) in different groups. Scale bar, 50 μM. The graph on the right showed SIRT3 fluorescence intensity per nucleus, and the values of Sham + VEH group were normalized to 100% (n = 9). (E, F) Representative images of double immunofluorescence labeling of PGC1α and LepR at the bone marrow area of proximal tibia. Scale bar, 20 μM. The graph on the right showed PGC1α fluorescence intensity per LepR+ cells, and the values of Sham + VEH group were normalized to 100% (n = 9). Each value is mean ± SD. Significance was calculated using a one-way ANOVA followed by the Tukey’s post-hoc test (B, D and F). *p < 0.05 and **p < 0.01 compared to the Sham + VEH group. #p < 0.05 and ##p < 0.01 compared to the OVX + VEH group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For peroxisome proliferator-activated receptor gamma coactivator one alpha (PGC1α) and leptin receptor (LepR) immunofluorescence staining, sections were blocked with 10% goat serum (Gibco) and then incubated with primary antibodies (anti- PGC1α, 1:50, Proteintech; anti- M. Li et al. Redox Biology 61 (2023) 102649 LepR, 1:50, Invitrogen) at 4 ◦C overnight and with secondary antibodies for 1 h at 37 ◦C.

Techniques: Marker, Expressing, Immunohistochemical staining, Staining, Immunofluorescence, Fluorescence, Labeling