Journal: Frontiers in Neuroscience

Article Title: Regulation of neurons in the dorsal motor nucleus of the vagus by SIRT1

doi: 10.3389/fnins.2013.00270

Figure Lengend Snippet: Resveratrol increased spontaneous excitatory neurotransmission through SIRT1 activation in neurons of the dorsal motor nucleus of the vagus (DMV). (A) Continuous whole-cell patch-clamp recordings of spontaneous EPSCs (sEPSCs) at holding potential of −60 mV before (upper trace) and after (lower trace) resveratrol application. (B) Combined data showing increased frequency of sEPSCs following resveratrol application. * Significance ( p < 0.05). (C) Cumulative event probability plot of inter-event interval distribution in the recording shown in (A) . (D) Mean group data showing that in the presence of a selective SIRT1 inhibitor EX527 resveratrol failed to increase sEPSC frequency in DMV neurons. (E) Combined data showing no significant change in amplitude of sEPSCs following resveratrol application in the presence of EX527.

Article Snippet: The SIRT1 activator resveratrol (1–500 μ M, Tocris Bioscience) and the selective SIRT1 inhibitor EX527 (500 nM, Tocris Bioscience) were dissolved in ethanol and diluted in aCSF (final concentration of ethanol <0.1% by volume).

Techniques: Activation Assay, Patch Clamp

Journal: Frontiers in Neuroscience

Article Title: Regulation of neurons in the dorsal motor nucleus of the vagus by SIRT1

doi: 10.3389/fnins.2013.00270

Figure Lengend Snippet: Schematic illustration of resveratrol action on presynaptic terminals . SIRT1 activation with resveratrol results in increased glutamate release from presynaptic terminals. This mechanism depends on PI3-kinase dependent closure of K ATP channels leading to depolarization and increased neurotransmitter release.

Article Snippet: The SIRT1 activator resveratrol (1–500 μ M, Tocris Bioscience) and the selective SIRT1 inhibitor EX527 (500 nM, Tocris Bioscience) were dissolved in ethanol and diluted in aCSF (final concentration of ethanol <0.1% by volume).

Techniques: Activation Assay

Journal: Lipids in Health and Disease

Article Title: CircLDLR acts as a sponge for miR-667-5p to regulate SIRT1 expression in non-alcoholic fatty liver disease

doi: 10.1186/s12944-022-01740-9

Figure Lengend Snippet: miR-667-5p can target SIRT1 to regulate the autophagy signaling pathway. A qRT-PCR detection of miR-667-5p level in Hepa1-6 cells. *** P < 0.001 vs. NC mimic group. B qRT-PCR detection of SIRT1 mRNA expression in Hepa1-6 cells. * P < 0.05 vs. NC mimic group. C SIRT1 levels in NAFLD mice were detected by western blot. *** P < 0.001 vs. NC mimic group. D The potential binding sequences between circLDLR and SIRT1 were predicted by TargetScan. E The binding association of miR-667-5p with SIRT1 was verified by determining the luciferase activity. * P < 0.05 vs. WT + NC mimic group. F and G The TG and TC contents in Hepa1-6 cells were measured via enzymatic method. * P < 0.05 vs. vector + NC mimic group; # P < 0.05 vs. vector + miR-667-5p mimic group. H SIRT1, p62, LC3, and mTOR expressions in Hepa1-6 cells were determined using western blot and relative quantification by densitometry. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. vector + NC mimic group; # P < 0.05, ### P < 0.001 vs. SIRT1 OE + NC mimic group; & P < 0.05, &&& P < 0.001 vs. vector + miR-667-5p mimic group

Article Snippet: The luciferase reporter vectors circLDLR-Wt, circLDLR-Mut, SIRT1-Wt, and SIRT1-Mut were created via inserting wild-type (Wt) or mutant (Mut) sequences of circLDLR or SIRT1 3ʹUTR with the putative miR_668_5p binding sites into the pmirGLO plasmid (Promega, Madison, WI, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Binding Assay, Luciferase, Activity Assay, Plasmid Preparation

Journal: Lipids in Health and Disease

Article Title: CircLDLR acts as a sponge for miR-667-5p to regulate SIRT1 expression in non-alcoholic fatty liver disease

doi: 10.1186/s12944-022-01740-9

Figure Lengend Snippet: circLDLR regulates the SIRT1-autophagy signaling pathway to alleviate cells’ lipid accumulation. A The SIRT1 mRNA expression in Hepa1-6 cells. *** P < 0.001 vs. NC siRNA group. B The SIRT1 protein level in Hepa1-6 cells was determined using western blot. *** P < 0.001 vs. NC siRNA group. C ORO staining was determined to study the lipogenesis of si-SIRT1 in Hepa1-6 cells. ** P < 0.01 vs. NC group. D and E The TG and TC contents in Hepa1-6 cells were detected via an enzymatic method. * P < 0.05 vs. NC group; # P < 0.05 vs. vector group. F LC3 and p62 expressions in Hepa1-6 cells were determined through western blot and relative quantification. * P < 0.05, *** P < 0.001 vs. vector group; ## P < 0.01, ### P < 0.001 vs. circLDLR OE group; && P < 0.01, &&& P < 0.001 vs. vector + si-SIRT1 group

Article Snippet: The luciferase reporter vectors circLDLR-Wt, circLDLR-Mut, SIRT1-Wt, and SIRT1-Mut were created via inserting wild-type (Wt) or mutant (Mut) sequences of circLDLR or SIRT1 3ʹUTR with the putative miR_668_5p binding sites into the pmirGLO plasmid (Promega, Madison, WI, USA).

Techniques: Expressing, Western Blot, Staining, Plasmid Preparation

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The circadian rhythmicity of Stat3 mRNA expression in adult rat pineal glands determined by qPCR. The normalized values are expressed as relative mRNA levels. The data indicated by dots were fitted with a cosine curve using the cosinor analysis method. Each point represents the mean ± SEM from five animals.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Expressing

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The activity of AANAT in organotypic cultures of pineal glands. The cultures were stimulated with STAT3 inhibitor WP1066 (WP; 5 μM) for 6 h, isoprenaline (ISO; 2 μM) for 4 h, or pretreated with WP1066 for 2 h followed by ISO for 4 h (ISO + WP). The data are the mean of 10–17 pineal glands from four independent experiments (numbers within the columns). The statistical significance is shown with unpaired two-tailed t test.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Activity Assay, Two Tailed Test

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The changes in aa-nat expression ( A ), Icer mRNA ( C ), and Stat3 mRNA ( D ) in the rat pinealocytes. The insert in ( A ) indicates the first two column values in a different scale of the y-axis. ( B ) Representative western blots of STAT3 and its phosphorylated forms after transfection of pinealocytes with Stat3 siRNA (The original western blots are in the ). After 2 days of incubation with Stat3 siRNA, cells were treated with 2 µM ISO for 4 h. Data are presented as the means ± SEM of four to nine experiments (different dissections; numbers within the columns). The statistical significance is shown with unpaired two-tailed t test.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Expressing, Western Blot, Transfection, Incubation, Two Tailed Test

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The activity of AANAT in adult rat pineal glands. Animals were treated with STAT3 inhibitors, WP1066 (WP; 40 mg/kg) or AZD1480 (AZD; 40 mg/kg), or a combination of inhibitors with 1 min light pulse (LP). Controls (C) were treated with vehiculum. Animals were injected either during the subjective day at CT4 ( A ) or during the subjective night CT15 ( B ) and euthanized 5 h later. For better clarity, there is a difference in the y-axis scale between ( A , B ). The data are the mean of four to seven pineal glands (numbers within the columns). The statistical significance is shown with unpaired two-tailed t test.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Activity Assay, Injection, Two Tailed Test

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: LPS-induced changes in Stat3 gene expression in rat pineal glands. Adult rats were injected with LPS (1 mg/kg) during the subjective day at CT8, pineal glands were sampled 4 h later and then in 3-h intervals throughout the 24-h cycle. The normalized values are expressed as relative mRNA levels. * indicates the p < 0.001 of multiple t -tests with the Sidak–Bonferroni post hoc test. Each point represents the mean ± SEM from four animals.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Gene Expression, Injection

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The activity of AANAT in adult rat pineal glands ( A ) and in organotypic cultures of pineal glands ( B ). At CT15, four adult males were treated with LPS (1 mg/kg) or with STAT3 inhibitors WP1066 (40 mg/kg) for 1 h followed by the LPS for 4 h. The pineal glands were sampled 5 h later. The organotypic cultures were stimulated with LPS (10 µg/mL) for 5 h, isoprenaline (ISO; 2 μM) for 4 h, a combination of both (ISO + LPS), or were pre-treated with STAT3 inhibitor WP1066 (WP; 5 μM) for 1 h. The data are the mean of 8 to 21 pineal glands from four independent experiments (numbers within the columns). The statistical significance is shown with unpaired two-tailed t test.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Activity Assay, Two Tailed Test

Journal: Life

Article Title: The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

doi: 10.3390/life11101105

Figure Lengend Snippet: The changes in aa-nat expression ( A ), Icer mRNA ( C ), and Stat3 mRNA ( D ) in the rat pinealocytes. The insert in ( A ) indicates the values of the first three columns with a difference in the y-axis scale. ( B ) Representative western blots of TLR4 receptor in three randomly selected pinealocytes cultures. The original western blots are in the . After 2 days of incubation with Stat3 siRNA, cells were treated with LPS (10 µg/mL) for 5 h, ISO (2 µM) for 4 h, or with LPS for 1 h followed by ISO for 4 h. Data are presented as the means ± SEM of four to eight experiments (different dissections; numbers within the columns). The statistical significance is shown with unpaired two-tailed t test.

Article Snippet: On the day of experiment, the animals were released into DD and separated into nine groups; three groups received intraperitoneal injection of cell-permeable STAT3 inhibitor WP1066 (A11795, AdooQ Bioscience; 40 mg/kg [ ], or selective Janus kinases inhibitor shown to inhibit endogenous, as well as cytokine-induced, STAT3 activation AZD1480 (A10110, AdooQ Bioscience; 40 mg/kg [ ], or saline.

Techniques: Expressing, Western Blot, Incubation, Two Tailed Test

Journal: PLOS ONE

Article Title: Ruscus aculeatus extract promotes RNase 7 expression through ERK activation following inhibition of late-phase autophagy in primary human keratinocytes

doi: 10.1371/journal.pone.0314873

Figure Lengend Snippet: ( A–G ) NHEKs were treated with RAE (0.1% [v/v]) for 72 h in the presence of wortmannin (10 μM), a PI3K inhibitor (A) or EX-527 (30 μM), a SIRT1 inhibitor (B–G). ( A, B ) mRNA expression levels of RNase 7 were determined by RT-qPCR. ( C–G ) Protein expression levels of RNase 7 (C), phosphorylated ERK (D), LC3-Ⅱ (E), p62 (F), and phosphorylated S6 (G) were analysed by western blotting. ( A–G ) Data represent the mean ± SD of three independent experiments. ( A–F ) Different letters indicate significant differences based on post-hoc Tukey’s test results. P < 0.05. ( G ) Differences in results of control versus RAE, control versus EX, and control versus RAE + EX were analysed by Bonferroni correction. # P < 0.0166 (0.05/3). EX: EX-527.

Article Snippet: Hydroxychloroquine sulfate (HCQ), bafilomycin A1 (BA1), PD98059, wortmannin, EX-527, and resveratrol were purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control