Journal: Current Research in Food Science

Article Title: Dietary kaempferol exerts anti-obesity effects by inducing the browing of white adipocytes via the AMPK/SIRT1/PGC-1α signaling pathway

doi: 10.1016/j.crfs.2024.100728

Figure Lengend Snippet: KPF increased energy expenditure and induced white adipose tissue Browning via AMPK/SIRT1/PGC-1α pathway. (A, B, C and D) Ex vivo oxygen consumption of iWAT and eWAT homogenates from NCD (A, B) or HFD (C, D) mice. (E) Immunoblot of the cell related protein in iWAT from mice on NCD. (G) Immunoblot of the cell related protein in iWAT from mice on HFD. Specific bands were quantified and are presented as graphs (F, H). Data shown are mean ± SD (n = 3–6). *p < 0.05 , **p < 0.01, vs. Control.

Article Snippet: Rabbit antibodies for β-actin (93473), AMPK (2532), p-AMPK (50081), PGC-1α (2178), PPARγ (2435), and Mouse antibodies for SIRT1 (8469) were obtained from Cell Signaling Technologies (Danvers, MA), while Rabbit antibodies for UCP1 (ab234430), PRDM16 (ab191838) was procured from Abcam (Cambridge, United Kingdom).

Techniques: Ex Vivo, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Hsp90 Stabilizes SIRT1 Orthologs in Mammalian Cells and C. elegans

doi: 10.3390/ijms19113661

Figure Lengend Snippet: Hsp90 inhibition depletes SIRT1 protein in COS-7 and HepG2 cells. ( a ) Effect of GA on SIRT1 and Raf-1 protein levels. Western blots of lysates from COS-7 cells treated by GA for 20 h. Images are representatives of three experiments for SIRT1 and two experiments for Raf-1. ( b ) Quantification of protein levels from the experiment shown in panel ( a ). Values are means ± S.D. of three (SIRT1) or two (Raf-1) experiments and were statistically compared with the respective untreated controls. * p < 0.05, ** p < 0.01 by two-tailed unpaired t -test. ( c ) Timescale of GA treatment on SIRT1 and Raf-1 protein levels. Western blots of lysates from COS-7 cells treated by 1 μg/mL GA for the indicated times. Images are representatives of two experiments. C: 48 h vehicle control. ( d ) Effect of GA on SIRT2 protein level. Western blots of lysates from COS-7 cells treated with GA for 20 h. Images are representatives of two experiments. ( e ) Effect of GA on SIRT1 protein level in human hepatoma cells. Western blots of lysates from HepG2 cells treated by GA for 48 h. Images are representatives of two experiments.

Article Snippet: Rabbit polyclonal antibodies against SIRT1 and SIRT2 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Inhibition, Western Blot, Two Tailed Test, Control

Journal: International Journal of Molecular Sciences

Article Title: Hsp90 Stabilizes SIRT1 Orthologs in Mammalian Cells and C. elegans

doi: 10.3390/ijms19113661

Figure Lengend Snippet: Disruption of the Hsp90-SIRT1 interaction leads to destabilization and proteasomal degradation of SIRT1. ( a ) SIRT1 physically interacts with Hsp90 in a GA-sensitive manner. Western blots showing the co-precipitation of Hsp90 with SIRT1 from COS-7 cells treated by 1 μg/mL GA or vehicle for 3 h. pG, protein G control; C, control (DMSO vehicle). ( b ) GA induces destabilization and proteasomal degradation of SIRT1. Western blots of total cell lysates from cells treated by 1 μg/mL GA and/or 5 μM MG132 or DMSO vehicle for 20 h. Images are representatives of two experiments.

Article Snippet: Rabbit polyclonal antibodies against SIRT1 and SIRT2 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Disruption, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Hsp90 Stabilizes SIRT1 Orthologs in Mammalian Cells and C. elegans

doi: 10.3390/ijms19113661

Figure Lengend Snippet: A conserved chaperone-client interaction between Hsp90 and SIRT1 orthologs. ( a ) SIRT1 orthologs possess extensive N- and C-terminal domains. Domain structures of yeast, worm, and human SIRT1 orthologs and the SIRT1 paralog SIRT2. Numbers denote the number of amino acid residues at the respective domain boundaries. The catalytic core domains (CD) are in light blue, N- and C-terminal regions are in white. ( b ) Hypothetical model of the conformational regulation of SIRT1 (and respective orthologs) by Hsp90. Hsp90 binds the nascent, unstable SIRT1 in the cytosol and stabilizes it in a probably open, partially unfolded conformation. Hsp90 binding facilitates various interactions which ensure activation by signals, substrate, and co-factor binding. Active, closed SIRT1 might dissociate from, or might remain in loose complex with, Hsp90 and exerts its cellular functions. Reduction in Hsp90 capacity, by pharmacological inhibition, genetic down-regulation or stresses, induces SIRT1 destabilization and proteasomal degradation, which in turn attenuates SIRT1 mediated processes.

Article Snippet: Rabbit polyclonal antibodies against SIRT1 and SIRT2 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Binding Assay, Activation Assay, Inhibition

Journal: Experimental and therapeutic medicine

Article Title: MicroRNA-34a regulates proliferation and apoptosis of gastric cancer cells by targeting silent information regulator 1.

doi: 10.3892/etm.2018.5920

Figure Lengend Snippet: Figure 4. Gel images of PCR products of recombinant vectors. Following PCR, the double enzyme restriction digestion and agarose gel electrophoresis was performed to determine the fragment size. (A) Enzyme‑digested SIRT1‑3'‑UTR fragments. (B) Lane 1, ddH2O; lane 2, negative control (empty vector self‑ligation); lane 3, positive control (GAPDH); lane 4, MW scale (molecular weight of marker protein); lanes 5‑12, recombinant SIRT1‑3'‑UTR luciferase reporter vector plasmid. PCR, polymerase chain reaction; miR‑34a, microRNA‑34a; SIRT1, sirtuin 1; UTR, untranslated region.

Article Snippet: Rabbit anti-human SIRT1 antibody was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA), GAPDH antibody (cat no. AF0006), SIRT1 antibody (cat no. AF1267), horseradish peroxide conjugated goat-anti-rabbit Immunoglobulin g (H+L) secondary antibody (cat no. A0208) and the ECL kit were purchased from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Recombinant, Agarose Gel Electrophoresis, Negative Control, Plasmid Preparation, Positive Control, Molecular Weight, Marker, Luciferase, Polymerase Chain Reaction

Journal: Experimental and therapeutic medicine

Article Title: MicroRNA-34a regulates proliferation and apoptosis of gastric cancer cells by targeting silent information regulator 1.

doi: 10.3892/etm.2018.5920

Figure Lengend Snippet: Figure 3. Results of miR‑34a targeting gene prediction. (A) Binding sites for miR‑34a within the SIRT1‑3'‑UTR extend from 891‑897 and 1434‑1440 bp. (B) Seeding sequence of SIRT1‑3'‑UTR and species homology analysis derived from miRanda, TargetScan and PICTAR databases. miR‑34a, microRNA‑34a; SIRT1, sirtuin 1; UTR, untranslated region.

Article Snippet: Rabbit anti-human SIRT1 antibody was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA), GAPDH antibody (cat no. AF0006), SIRT1 antibody (cat no. AF1267), horseradish peroxide conjugated goat-anti-rabbit Immunoglobulin g (H+L) secondary antibody (cat no. A0208) and the ECL kit were purchased from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Binding Assay, Sequencing, Derivative Assay

Journal: Experimental and therapeutic medicine

Article Title: MicroRNA-34a regulates proliferation and apoptosis of gastric cancer cells by targeting silent information regulator 1.

doi: 10.3892/etm.2018.5920

Figure Lengend Snippet: Figure 7. Relative luciferase activity of each group. Group 1, 3'‑UTR empty plasmid + miRNA empty plasmid; group 2, 3'‑UTR empty plasmid + miR‑34a over‑expression plasmid; group 3, SIRT1‑3'‑UTR recombinant plasmid + miRNA empty plasmid; group 4, SIRT1‑3'‑UTR recombinant plasmid + miR‑34a overexpression plasmid; group 5, SIRT1‑3'‑UTR mutant plasmid + miRNA empty plasmid; group 6, SIRT1‑3'‑UTR mutant plasmid + miR‑34a over‑expression plasmid. Values are expressed as the mean ± standard deviation for three experiments. **P<0.01. miR‑34a, microRNA‑34a; SIRT1, sirtuin 1; UTR, untranslated region.

Article Snippet: Rabbit anti-human SIRT1 antibody was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA), GAPDH antibody (cat no. AF0006), SIRT1 antibody (cat no. AF1267), horseradish peroxide conjugated goat-anti-rabbit Immunoglobulin g (H+L) secondary antibody (cat no. A0208) and the ECL kit were purchased from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Luciferase, Activity Assay, Plasmid Preparation, Recombinant, Over Expression, Mutagenesis, Standard Deviation

Journal: Experimental and therapeutic medicine

Article Title: MicroRNA-34a regulates proliferation and apoptosis of gastric cancer cells by targeting silent information regulator 1.

doi: 10.3892/etm.2018.5920

Figure Lengend Snippet: Figure 9. Relative expression of SIRT1 mRNA. The groups were as follows: miR‑34a, infected hsa‑miR‑34a over‑expression viral group; CON, unin fected non‑viral group; NC, infected negative control viral group. Values are expressed as the mean ± standard deviation. The experiment was performed in triplicate. miR‑34a, microRNA‑34a; SIRT1, sirtuin 1.

Article Snippet: Rabbit anti-human SIRT1 antibody was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA), GAPDH antibody (cat no. AF0006), SIRT1 antibody (cat no. AF1267), horseradish peroxide conjugated goat-anti-rabbit Immunoglobulin g (H+L) secondary antibody (cat no. A0208) and the ECL kit were purchased from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Expressing, Infection, Negative Control, Standard Deviation

Journal: Experimental and therapeutic medicine

Article Title: MicroRNA-34a regulates proliferation and apoptosis of gastric cancer cells by targeting silent information regulator 1.

doi: 10.3892/etm.2018.5920

Figure Lengend Snippet: Figure 10. Relative expression of SIRT1 protein determined by western blot analysis. (A) Quantified western blot results. (B) Images of western blot analysis. The groups were as follows: miR‑34a, infected hsa‑miR‑34a over‑expression viral group; CON, uninfected non‑viral group; NC, infected negative control viral group. Values are expressed as the mean ± standard deviation for three experiments. **P<0.01 vs. NC group, ##P<0.01 vs. CON group. miR‑34a, microRNA‑34a; SIRT1, sirtuin 1.

Article Snippet: Rabbit anti-human SIRT1 antibody was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA), GAPDH antibody (cat no. AF0006), SIRT1 antibody (cat no. AF1267), horseradish peroxide conjugated goat-anti-rabbit Immunoglobulin g (H+L) secondary antibody (cat no. A0208) and the ECL kit were purchased from Beyotime Institute of Biotechnology (Haimen, China).

Techniques: Expressing, Western Blot, Infection, Negative Control, Standard Deviation