Journal: RSC Chemical Biology

Article Title: Mechanism-based inhibitors of SIRT2: structure–activity relationship, X-ray structures, target engagement, regulation of α-tubulin acetylation and inhibition of breast cancer cell migration

doi: 10.1039/D0CB00036A

Figure Lengend Snippet: Scheme 1 Structure–activity relationship of SIRT2 inhibitors. Potencies for inhibition of the deacylase activity of recombinant SIRT2 (100 nM) against QPKKac (shown with asterisks**) or ETDKmyr (shown in bold/italics) as substrate (50 mM) are given as mean IC50 values standard deviation (SD) or %inhibition. (a) Lead compounds. (b) Amide isosteres. (c) Optimization of position i + 2. (d) Optimization of position + 1. (e) Optimization of N-terminal group. (f) Final inhibitors. Data are based on two individual experiments performed in duplicate. See the ESI† (Scheme S1, Fig. S1–S3 and Table S2) for dose–response curves and selectivity profiling of additional inhibitors (S17–S23) against SIRT1–3. *A single asterisk denotes a stereogenic center.

Article Snippet: For general experimental information, chemical synthesis, compound characterization data, additional methods, and copies of analytical HPLC chromatograms and NMR spectra of novel compounds, please consult the ESI.† Materials and methods for the biochemical assays SIRT1 (aa 193–741 with N-terminal GST-tag, Z60% purity; cat. #50012), SIRT2 (aa 50–356 with C-terminal His-tag, Z90% purity; cat. #50013), SIRT3 (aa 102–399 with N-terminal GST-tag; Z64% purity; cat. #50014), and SIRT6 (full length with N-terminal GST-tag, Z75% purity; cat. #50017) from BPS Biosciences (San Diego, CA).

Techniques: Activity Assay, Inhibition, Recombinant, Standard Deviation

Journal: RSC Chemical Biology

Article Title: Mechanism-based inhibitors of SIRT2: structure–activity relationship, X-ray structures, target engagement, regulation of α-tubulin acetylation and inhibition of breast cancer cell migration

doi: 10.1039/D0CB00036A

Figure Lengend Snippet: Fig. 4 Pre-incubation and HPLC-based sirtuin assays. (a) Inhibition of SIRT1 (20 nM) and SIRT2 (20 nM)-mediated deacetylation of the H3K9ac residue in a dodecameric peptide (50 mM) determined by HPLC. Error bars represent mean SEM of at least 2 independent experiments. See Table S4 (ESI†) for IC50 values against sirtuin mediated H3K9ac deacetylation. (b) Pre-incubation (30 min), with or without NAD+ (500 mM) for selected compounds against SIRT2 (100 nM) demyristoylation using ETDKmyr (50 mM) as substrate. Mean SEM based on at least 2 independent experiments performed in duplicate.

Article Snippet: For general experimental information, chemical synthesis, compound characterization data, additional methods, and copies of analytical HPLC chromatograms and NMR spectra of novel compounds, please consult the ESI.† Materials and methods for the biochemical assays SIRT1 (aa 193–741 with N-terminal GST-tag, Z60% purity; cat. #50012), SIRT2 (aa 50–356 with C-terminal His-tag, Z90% purity; cat. #50013), SIRT3 (aa 102–399 with N-terminal GST-tag; Z64% purity; cat. #50014), and SIRT6 (full length with N-terminal GST-tag, Z75% purity; cat. #50017) from BPS Biosciences (San Diego, CA).

Techniques: Incubation, Inhibition, Residue

Journal: RSC Chemical Biology

Article Title: Mechanism-based inhibitors of SIRT2: structure–activity relationship, X-ray structures, target engagement, regulation of α-tubulin acetylation and inhibition of breast cancer cell migration

doi: 10.1039/D0CB00036A

Figure Lengend Snippet: Fig. 6 Cellular target engagement and SIRT2i effect on perinuclear a-tubulin acetylation. (a) Cellular thermal shift of SIRT2 in HEK293T cells subjected to 1 h treatment with inhibitor at designated concentrations or DMSO (vehicle). Please see Fig. S8 (ESI†) for complete dataset against SIRT1–3 (n = 3). (b–g) Immunofluorescence images (40) of MCF-7 cells subjected to 6 h treatment with inhibitor [TSA (5 mM), 26 (5 mM), TM (25 mM), 26-D (25 mM)] or DMSO (vehicle). (g) Zoomed image for compound 26. DAPI (blue, nuclear counter- stain) and Ac-a-tubulin (green). For additional images (single-filtered and zoomed) for all tested compounds, please see Fig. S9 (ESI†). The data are representative images from two individual experiments.

Article Snippet: For general experimental information, chemical synthesis, compound characterization data, additional methods, and copies of analytical HPLC chromatograms and NMR spectra of novel compounds, please consult the ESI.† Materials and methods for the biochemical assays SIRT1 (aa 193–741 with N-terminal GST-tag, Z60% purity; cat. #50012), SIRT2 (aa 50–356 with C-terminal His-tag, Z90% purity; cat. #50013), SIRT3 (aa 102–399 with N-terminal GST-tag; Z64% purity; cat. #50014), and SIRT6 (full length with N-terminal GST-tag, Z75% purity; cat. #50017) from BPS Biosciences (San Diego, CA).

Techniques: Drug discovery, Immunofluorescence, Staining

Journal: Antioxidants

Article Title: Pterostilbene, a Resveratrol Derivative, Improves Ovary Function by Upregulating Antioxidant Defenses in the Aging Chickens via Increased SIRT1/Nrf2 Expression

doi: 10.3390/antiox13080935

Figure Lengend Snippet: Primers sequences for PCR analysis.

Article Snippet: Before treatment with PTS (0.5 μM) and D-gal (200 mM), cultured SWFs underwent a 24 h pretreatment with 20 μM EX-527, a sirtuin1 (SIRT1) inhibitor (49843-98-3, MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Sequencing

Journal: Antioxidants

Article Title: Pterostilbene, a Resveratrol Derivative, Improves Ovary Function by Upregulating Antioxidant Defenses in the Aging Chickens via Increased SIRT1/Nrf2 Expression

doi: 10.3390/antiox13080935

Figure Lengend Snippet: The suppression of EX-527 on the promotion of cell antioxidant ability and inhibition of cell apoptosis by PTS. ( A ) Effect of different treatments on the expression of apoptosis-related protein expression. Bax (A1) BCL-2 (A2), Caspase3 (A3). ( B ) Effect of different treatments on the expression of apoptosis-related genes expression. Bax (B1) Caspase3 (B2), BCL-2 (B3). ( C ) Effect of different treatments on the expression of Nrf2, p-Nrf2 and SIRT1 protein expression. p-Nrf2/Nrf2 (C1), SIRT1 (C2). ( D ) Effect of different treatments on the expression of Nrf2 and SIRT1 genes expression. Nrf2 (D1), SIRT1 (D2). ( E ) Effect of different treatments on the expression of antioxidant-related genes expression. CAT (E1), SOD (E2), Mgst (E3), Gsta (E4). All gene and protein expression levels were analyzed with β-actin as a control. Significant differences between groups are shown by distinct lowercase letters in a test ( p < 0.05).

Article Snippet: Before treatment with PTS (0.5 μM) and D-gal (200 mM), cultured SWFs underwent a 24 h pretreatment with 20 μM EX-527, a sirtuin1 (SIRT1) inhibitor (49843-98-3, MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Inhibition, Expressing, Control

Journal: Aging (Albany NY)

Article Title: The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model

doi: 10.18632/aging.203237

Figure Lengend Snippet: CR and the AMPK-SIRT1-mTOR network. AMPKs were analyzed by qPCR (mRNA levels) and Western Blot (protein levels) in pectoralis major (PM) ( A , B ) and quadriceps femoris vastus medialis ( QF - VM ) ( C , D ). SIRT1 was analyzed in PM and QF- VM ( E ) with the Mouse NAD-Dependent Deacetylase Sirtuin-1 (SIRT1) ELISA Kit (CUSABIO) for quantitative determination, according to the assay kit protocol . Values are expressed in ng/ml. mTOR mRNA in PM and QF- VM ( F ). Immunoblot results and protein expression of mTOR ( G ) and mTOR-S2448 ( H ) in PM and QF- VM . For the qPCR assay, each primer was analyzed with SYBR Green fluorescence detection and the transcript levels, expressed as a %, were normalized to those of the endogenous control 18s rRNA. Protein expressions were obtained by Western Blot analysis and quantified with Image Lab 6.1 software. SD expression was set as 1 and the relative protein levels were normalized as a ratio of GAPDH expression. The data are the mean ± s.d. *P < 0.05; **P < 0.01 vs SD, unless otherwise specified.

Article Snippet: One hundred mg of PM and QF- VM tissue homogenates from the SD (n=10), SD-CR (n=19), HC (n=14), and HC-CR (n=33) were analyzed with a Mouse NAD-Dependent Deacetylase Sirtuin-1 (SIRT1) ELISA Kit (CUSABIO) for the quantitative determination of SIRT1, according to its manual instructions (Supplementary Material 3).

Techniques: Western Blot, Histone Deacetylase Assay, Enzyme-linked Immunosorbent Assay, Expressing, SYBR Green Assay, Fluorescence, Control, Software

Journal: Cell Death & Disease

Article Title: The NR3C2-SIRT1 signaling axis promotes autophagy and inhibits epithelial mesenchymal transition in colorectal cancer

doi: 10.1038/s41419-025-07575-3

Figure Lengend Snippet: A Volcano plot of downstream regulatory gene after NR3C2 overexpression. B TIMER prediction of the relationship between NR3C2 and SIRT1. left: COAD; right: READ; C Western blot detection of SIRT1 expression after NR3C2 overexpression and NR3C2 knockdown, * P < 0.05, ** P < 0.01; D Fluorescence double staining to detect the expression of NR3C2 and SIRT1 (Blue: DAPI; Red: NR3C2; Green: SIRT1). Upper: HCT116, lower: SW620; E ChIP assay to detect the relationship between NR3C2 and SIRT1. * P < 0.05. F Transwell migration experiment validating that siSIRT1 increases the decrease in cells migration. Upper: HCT116, lower: RKO, right: statistical chart, * P < 0.05(HCT116), # P < 0.05(RKO).

Article Snippet: Cell lysates were incubated with antibodies (anti-SIRT1 antibody or anti-NR3C2 antibody or control IgG), followed by incubation with protein A/G magnetic beads (MCE, USA) at 4 °C overnight.

Techniques: Over Expression, Western Blot, Expressing, Knockdown, Fluorescence, Double Staining, Migration

Journal: Cell Death & Disease

Article Title: The NR3C2-SIRT1 signaling axis promotes autophagy and inhibits epithelial mesenchymal transition in colorectal cancer

doi: 10.1038/s41419-025-07575-3

Figure Lengend Snippet: A Photograph of mouse lung metastasis model using RKO cells, right: weight change of mice; B HE staining of mouse lung tissues for RKO-vector and RKO-NR3C2; C Photograph of mouse lung metastasis model using HCT116 cells, right: weight change of mice; D HE staining of mouse lung tissues for HCT116-vector and HCT116-NR3C2; E Western blot detection of NR3C2, SIRT1, and P62 expression in the lung tissues of mice in the RKO group, * P < 0.05; F Western blot detection of NR3C2, SIRT1, and P62 expression in the lung tissues of mice in the HCT116 group, * P < 0.05; G IHC detection of NR3C2 expression in mouse lung tissues (50X and 400X); H IHC detection of SIRT1 expression in mouse lung tissues(50X and 400X).

Article Snippet: Cell lysates were incubated with antibodies (anti-SIRT1 antibody or anti-NR3C2 antibody or control IgG), followed by incubation with protein A/G magnetic beads (MCE, USA) at 4 °C overnight.

Techniques: Staining, Plasmid Preparation, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: The NR3C2-SIRT1 signaling axis promotes autophagy and inhibits epithelial mesenchymal transition in colorectal cancer

doi: 10.1038/s41419-025-07575-3

Figure Lengend Snippet: NR3C2 and SIRT1 constitute a pivotal signaling axis that modulates EMT in CRC cells through the intricate mechanism of autophagy.

Article Snippet: Cell lysates were incubated with antibodies (anti-SIRT1 antibody or anti-NR3C2 antibody or control IgG), followed by incubation with protein A/G magnetic beads (MCE, USA) at 4 °C overnight.

Techniques: