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Santa Cruz Biotechnology mouse sirt2 short hairpin rna lentiviral particles
<t>Sirt2</t> expression in regulatory T cells (Tregs) and Treg/T helper 17 (Th17) cells. (A) Representative flow cytometry zebra plots showing Foxp3‐tdTomato + interleukin (IL)‐17‐EGFP − Tregs and Foxp3‐tdTomato + IL‐17‐EGFP + Treg/Th17 in the spleens (SP), mesenteric lymph nodes (mLN), and lamina propria (LP). (B) The frequencies of IL‐17‐EGFP + Treg/Th17 cells in indicated tissues. (C) Sirt2 messenger RNA levels in IL‐17‐EGFP − Tregs and IL‐17−EGFP + Treg/Th17 cells that were sorted from colitic mice. (D) Sirt2 protein levels in IL‐17‐EGFP − Tregs and IL‐17‐EGFP + Treg/Th17 cells that were sorted from colitic mice. N = five to six per group. ** p < .01; *** p < .001. Student's t test. DSS, dextran sulfate sodium‐treated colitic mice; IL‐17 − , IL‐17‐EGFP − Tregs; IL‐17 + , IL‐17‐EGFP + Treg/Th17 cells; V, vehicle‐treated mice.
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1) Product Images from "Sirtuin2 suppresses the polarization of regulatory T cells toward T helper 17 cells through repressing the expression of signal transducer and activator of transcription 3 in a mouse colitis model"

Article Title: Sirtuin2 suppresses the polarization of regulatory T cells toward T helper 17 cells through repressing the expression of signal transducer and activator of transcription 3 in a mouse colitis model

Journal: Immunity, Inflammation and Disease

doi: 10.1002/iid3.1160

Sirt2 expression in regulatory T cells (Tregs) and Treg/T helper 17 (Th17) cells. (A) Representative flow cytometry zebra plots showing Foxp3‐tdTomato + interleukin (IL)‐17‐EGFP − Tregs and Foxp3‐tdTomato + IL‐17‐EGFP + Treg/Th17 in the spleens (SP), mesenteric lymph nodes (mLN), and lamina propria (LP). (B) The frequencies of IL‐17‐EGFP + Treg/Th17 cells in indicated tissues. (C) Sirt2 messenger RNA levels in IL‐17‐EGFP − Tregs and IL‐17−EGFP + Treg/Th17 cells that were sorted from colitic mice. (D) Sirt2 protein levels in IL‐17‐EGFP − Tregs and IL‐17‐EGFP + Treg/Th17 cells that were sorted from colitic mice. N = five to six per group. ** p < .01; *** p < .001. Student's t test. DSS, dextran sulfate sodium‐treated colitic mice; IL‐17 − , IL‐17‐EGFP − Tregs; IL‐17 + , IL‐17‐EGFP + Treg/Th17 cells; V, vehicle‐treated mice.
Figure Legend Snippet: Sirt2 expression in regulatory T cells (Tregs) and Treg/T helper 17 (Th17) cells. (A) Representative flow cytometry zebra plots showing Foxp3‐tdTomato + interleukin (IL)‐17‐EGFP − Tregs and Foxp3‐tdTomato + IL‐17‐EGFP + Treg/Th17 in the spleens (SP), mesenteric lymph nodes (mLN), and lamina propria (LP). (B) The frequencies of IL‐17‐EGFP + Treg/Th17 cells in indicated tissues. (C) Sirt2 messenger RNA levels in IL‐17‐EGFP − Tregs and IL‐17−EGFP + Treg/Th17 cells that were sorted from colitic mice. (D) Sirt2 protein levels in IL‐17‐EGFP − Tregs and IL‐17‐EGFP + Treg/Th17 cells that were sorted from colitic mice. N = five to six per group. ** p < .01; *** p < .001. Student's t test. DSS, dextran sulfate sodium‐treated colitic mice; IL‐17 − , IL‐17‐EGFP − Tregs; IL‐17 + , IL‐17‐EGFP + Treg/Th17 cells; V, vehicle‐treated mice.

Techniques Used: Expressing, Flow Cytometry

The effect of Sirt2 silencing on regulatory T cell (Treg)/T helper 17 (Th17) generation without stimulation. (A) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐EGFP and Foxp3‐tdTomato in sorted lamina propria CD4 + tdTomato + EGFP − Tregs after lentiviral transduction. 24 h, 24 h after transduction; 48 h, 48 h after transduction; Ctrl, control lentivirus encoding a scrambled siRNA; Fresh, freshly sorted Tregs; Sirt2 siRNA, Sirt2 siRNA‐encoding lentivirus. (B and C) The frequencies of Foxp3‐tdTomato + , Foxp3‐tdTomato − , and IL‐17‐EGFP + cells in Tregs 24 h (B) or 48 h (C) after lentiviral transduction. Ctrl, cells transfected with control lentivirus; Si‐S, cells transfected with Sirt2 siRNA‐encoding lentivirus. (D) Messenger RNA (mRNA) levels of indicated cytokines 48 h after lentiviral transduction. Fresh: freshly sorted Tregs. (E) Sirt2 mRNA levels after transduction. (F) Representative histograms showing the dilution of CellTrace Violet in activated conventional CD4 + T cells after 5‐day coculture with Tregs. Alone: conventional CD4 + T cells alone. Ctrl Tregs: conventional CD4 + T cells co‐cultured with control Tregs. Sirt2‐silenced Tregs: conventional CD4 + T cells co‐cultured with Sirt2‐silenced Tregs. (G) Statistics of CellTrace Violet intensities. N = three to six per group. * p < .05; ** p < .01; *** p < .001. Student's t test for (B), (C), and (E). One‐way analysis of variance for (D) and (G).
Figure Legend Snippet: The effect of Sirt2 silencing on regulatory T cell (Treg)/T helper 17 (Th17) generation without stimulation. (A) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐EGFP and Foxp3‐tdTomato in sorted lamina propria CD4 + tdTomato + EGFP − Tregs after lentiviral transduction. 24 h, 24 h after transduction; 48 h, 48 h after transduction; Ctrl, control lentivirus encoding a scrambled siRNA; Fresh, freshly sorted Tregs; Sirt2 siRNA, Sirt2 siRNA‐encoding lentivirus. (B and C) The frequencies of Foxp3‐tdTomato + , Foxp3‐tdTomato − , and IL‐17‐EGFP + cells in Tregs 24 h (B) or 48 h (C) after lentiviral transduction. Ctrl, cells transfected with control lentivirus; Si‐S, cells transfected with Sirt2 siRNA‐encoding lentivirus. (D) Messenger RNA (mRNA) levels of indicated cytokines 48 h after lentiviral transduction. Fresh: freshly sorted Tregs. (E) Sirt2 mRNA levels after transduction. (F) Representative histograms showing the dilution of CellTrace Violet in activated conventional CD4 + T cells after 5‐day coculture with Tregs. Alone: conventional CD4 + T cells alone. Ctrl Tregs: conventional CD4 + T cells co‐cultured with control Tregs. Sirt2‐silenced Tregs: conventional CD4 + T cells co‐cultured with Sirt2‐silenced Tregs. (G) Statistics of CellTrace Violet intensities. N = three to six per group. * p < .05; ** p < .01; *** p < .001. Student's t test for (B), (C), and (E). One‐way analysis of variance for (D) and (G).

Techniques Used: Flow Cytometry, Expressing, Transduction, Transfection, Cell Culture

The effect of Sirtuin2 (Sirt2) silencing on regulatory T cells (Tregs) after T helper 17 (Th17) induction. (A) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐enhanced green fluorescent protein (EGFP) and and Forkhead box P‐3 (Foxp3)‐tdTomato in sorted lamina propria CD4 + tdTomato + EGFP − Tregs after lentiviral transduction and Th17 induction. Ctrl, cells transduced with control lentivirus; Si‐S, cells transduced with Sirt2 siRNA‐encoding lentivirus. (B) The frequencies of IL‐17‐EGFP + cells and Foxp3‐tdTomato + cells in Tregs after lentiviral transduction and Th17 induction. (C, D) Messenger RNA levels of IL‐22 (C) and retinoic‐acid‐receptor‐related orphan nuclear receptor gamma (D) in Tregs after lentiviral transduction and Th17 induction. Fresh: freshly sorted Tregs as the negative control. (E) Representative flow cytometry histograms showing Ki67 stain in Tregs after lentiviral transduction and Th17 induction. The data represent two independent experiments. N = 6 per group. ** p < .01; *** p < .001. Student's t test.
Figure Legend Snippet: The effect of Sirtuin2 (Sirt2) silencing on regulatory T cells (Tregs) after T helper 17 (Th17) induction. (A) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐enhanced green fluorescent protein (EGFP) and and Forkhead box P‐3 (Foxp3)‐tdTomato in sorted lamina propria CD4 + tdTomato + EGFP − Tregs after lentiviral transduction and Th17 induction. Ctrl, cells transduced with control lentivirus; Si‐S, cells transduced with Sirt2 siRNA‐encoding lentivirus. (B) The frequencies of IL‐17‐EGFP + cells and Foxp3‐tdTomato + cells in Tregs after lentiviral transduction and Th17 induction. (C, D) Messenger RNA levels of IL‐22 (C) and retinoic‐acid‐receptor‐related orphan nuclear receptor gamma (D) in Tregs after lentiviral transduction and Th17 induction. Fresh: freshly sorted Tregs as the negative control. (E) Representative flow cytometry histograms showing Ki67 stain in Tregs after lentiviral transduction and Th17 induction. The data represent two independent experiments. N = 6 per group. ** p < .01; *** p < .001. Student's t test.

Techniques Used: Flow Cytometry, Expressing, Transduction, Negative Control, Staining

Involvement of the Sirtuin3 (Sirt3) signaling in Sirtuin2 (Sirt2)‐mediated effect. (A) Representative flow cytometry histograms showing the expression of phosphorylated Stat3 (left panel) and total Sta3 (right panel) in regulatory T cells (Tregs) after lentiviral transduction and T helper 17 (Th17) induction. Ctrl, cells transduced with control lentivirus; Isotype, isotype antibody control; Si‐S, cells transduced with Sirt2 siRNA‐encoding lentivirus. (B) Statistics of the mean fluorescent intensities of phosphorylated Stat3 (p‐Stat3) and total Stat3 in Tregs in (A). (C) The ratios of phosphorylated Stat3 (p‐Stat3) to total Stat3 in Tregs in (A). (D) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐EGFP and Foxp3‐tdTomato in Tregs after lentiviral transduction and Th17 induction in the presence or absence of C188‐9. V: DMSO. (E) Statistics of the frequencies of IL‐17‐EGFP + cells in (D). (F) Sirt2 messenger RNA levels in Tregs after lentiviral transduction and Th17 induction. (G) Immunoblotting images showing Stat3 proteins in Tregs on Day 0 (d0) and Day 3 (d3) after Th17 induction. (H) Statistics of normalized Stat3 protein levels in (G). N = five to six per group. ***: p < .001. Student's t test. DMSO, dimethyl sulfoxide.
Figure Legend Snippet: Involvement of the Sirtuin3 (Sirt3) signaling in Sirtuin2 (Sirt2)‐mediated effect. (A) Representative flow cytometry histograms showing the expression of phosphorylated Stat3 (left panel) and total Sta3 (right panel) in regulatory T cells (Tregs) after lentiviral transduction and T helper 17 (Th17) induction. Ctrl, cells transduced with control lentivirus; Isotype, isotype antibody control; Si‐S, cells transduced with Sirt2 siRNA‐encoding lentivirus. (B) Statistics of the mean fluorescent intensities of phosphorylated Stat3 (p‐Stat3) and total Stat3 in Tregs in (A). (C) The ratios of phosphorylated Stat3 (p‐Stat3) to total Stat3 in Tregs in (A). (D) Representative flow cytometry zebra plots showing the expression of interleukin (IL)‐17‐EGFP and Foxp3‐tdTomato in Tregs after lentiviral transduction and Th17 induction in the presence or absence of C188‐9. V: DMSO. (E) Statistics of the frequencies of IL‐17‐EGFP + cells in (D). (F) Sirt2 messenger RNA levels in Tregs after lentiviral transduction and Th17 induction. (G) Immunoblotting images showing Stat3 proteins in Tregs on Day 0 (d0) and Day 3 (d3) after Th17 induction. (H) Statistics of normalized Stat3 protein levels in (G). N = five to six per group. ***: p < .001. Student's t test. DMSO, dimethyl sulfoxide.

Techniques Used: Flow Cytometry, Expressing, Transduction, Western Blot

Overview of the findings. Foxp3, Forkhead box P‐3; Sirt2, Sirtuin2; Sirt3, Sirtuin3; RORγt, retinoic‐acid‐receptor‐related orphan nuclear receptor gamma T; Th17, T helper 17; Treg, regulatory T cell.
Figure Legend Snippet: Overview of the findings. Foxp3, Forkhead box P‐3; Sirt2, Sirtuin2; Sirt3, Sirtuin3; RORγt, retinoic‐acid‐receptor‐related orphan nuclear receptor gamma T; Th17, T helper 17; Treg, regulatory T cell.

Techniques Used:


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