ser9 5b3 cell signaling technology 9323p rrid ab 2115201 rabbit  (Cell Signaling Technology Inc)

Journal: Nature metabolism

Article Title: Fasting-induced FOXO4 blunts human CD4 + T helper cell responsiveness

doi: 10.1038/s42255-021-00356-0

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-STAT6 , Cell Signaling , Cat. #5397S; RRID:AB_11220421.

Techniques: Virus, Recombinant, Cell Stimulation, Staining, Flow Cytometry, CyQUANT Assay, Proliferation Assay, Enzyme-linked Immunosorbent Assay, Control, Software, Cell Isolation, In Vitro, Transfection

Journal: The Journal of Clinical Investigation

Article Title: MicroRNA-210 overexpression promotes psoriasis-like inflammation by inducing Th1 and Th17 cell differentiation

doi: 10.1172/JCI97426

Figure Lengend Snippet: (A) miR-210 target sequences (in red) of the 3′-UTR of STAT6 (A, upper panel) and LYN (A, lower panel) mRNAs and corresponding mutant sequences, which were included in luciferase reporter vectors. (B) Luciferase activities were measured in HEK293T cells cotransfected with luciferase reporter vectors and agomir-210 or agomir-NC, n = 3 per group. (C) Normal CD4+ T cells were transfected with agomir-210 or agomir-NC, and psoriatic CD4+ T cells were transfected with antagomir-210 or antagomir-NC. STAT6 (upper panel) and LYN (lower panel) protein levels were analyzed. (D) The protein levels of STAT6 and LYN in splenic CD4+ T cells from IMQ-treated WT and KO mice. (E–H) STAT6 and LYN protein levels in CD4+ T cells (E and F) and skin lesions (G and H) derived from psoriasis patients and IMQ-induced psoriasis-like mouse models (BALB/c) as well as their healthy controls. Data (B and C) are representative of at least 3 independent experiments. Data represent the mean ± SEM. *P < 0.05, **P < 0.01. NS, not significant. Two-tailed unpaired Student’s t test (B) was used.

Article Snippet: Rabbit anti–HIF-1α Ab (1:1,000; Cell Signal Technology, catalog 14179), rabbit anti-STAT6 Ab (1:1,000; Cell Signal Technology, catalog 9362), mouse anti-LYN Ab (1:1,000; Abcam, catalog ab1890), mouse anti–β-actin Ab (1:2,000, Abcam, catalog ab6276) or goat anti-GAPDH Ab (1:2,000, Abcam, catalog ab9483) was used.

Techniques: Mutagenesis, Luciferase, Transfection, Derivative Assay, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: MicroRNA-210 overexpression promotes psoriasis-like inflammation by inducing Th1 and Th17 cell differentiation

doi: 10.1172/JCI97426

Figure Lengend Snippet: (A and B) Normal human CD4+ T cells were transfected with STAT6 siRNAs and overexpression plasmid (A, n = 3) or LYN siRNAs and overexpression plasmid (B, n = 3) separately for 48 hours. STAT6 and LYN protein levels (left panel), and the mRNA levels of IL17A, IL17F, IFNG, and IL4 (middle and right panels) were detected in transfected cells. (C and D) Normal human CD4+ T cells were transfected with agomir-210 or agomir-NC, and after 24 hours, the STAT6 overexpression plasmid or plasmid control (C, n = 3) and LYN overexpression plasmid or plasmid control (D, n = 3) were transfected into cells. Then, cells were collected to detect the protein levels of STAT6 or LYN (left panel) and the mRNA levels of IL17A, IL17F, IFNG, and IL4 (right panel) at 48 hours after transfection. Similar results were obtained from 3 independent experiments. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant. Two-tailed unpaired Student’s t test (A and B) or 1-way ANOVA with Bonferroni’s post hoc test (C and D) was used.

Article Snippet: Rabbit anti–HIF-1α Ab (1:1,000; Cell Signal Technology, catalog 14179), rabbit anti-STAT6 Ab (1:1,000; Cell Signal Technology, catalog 9362), mouse anti-LYN Ab (1:1,000; Abcam, catalog ab1890), mouse anti–β-actin Ab (1:2,000, Abcam, catalog ab6276) or goat anti-GAPDH Ab (1:2,000, Abcam, catalog ab9483) was used.

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: MicroRNA-210 overexpression promotes psoriasis-like inflammation by inducing Th1 and Th17 cell differentiation

doi: 10.1172/JCI97426

Figure Lengend Snippet: (A and B) The expression of miR-210 in normal human CD4+ T cells stimulated with IL-23, TGF-β, IL-6, or IL-1β (A, n = 3 per group). The protein levels of HIF-1α, STAT6, and LYN in cells stimulated with IL-23 or TGF-β (B). (C) The miR-210 expression levels in human CD4+ T cells transfected with HIF-1α siRNA or siRNA controls followed by stimulation with IL-23 or TGF-β for 24 hours (n = 3 per group). (D) CD4+ T cells were transfected with HIF-1α overexpression plasmid (upper panel) or were stimulated with TGF-β (lower 2 panels). Co-IP and Western blot showed the interaction between HIF-1α and P300. (E–G) The enrichment of HIF-1α, P300, and H3ac on the miR-210 promoter in psoriatic CD4+ T cells transfected with HIF-1α siRNA or controls (E, n = 3), normal CD4+ T cells transfected with HIF-1α plasmid or controls (F, n = 3), or normal CD4+ T cells stimulated with TGF-β (G, n = 3). Data (B and D) are representative of at least 3 independent experiments. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. One-way ANOVA with Dunnett’s post hoc test (A and G) or 2-tailed unpaired Student’s t test (C, E, and F) was used.

Article Snippet: Rabbit anti–HIF-1α Ab (1:1,000; Cell Signal Technology, catalog 14179), rabbit anti-STAT6 Ab (1:1,000; Cell Signal Technology, catalog 9362), mouse anti-LYN Ab (1:1,000; Abcam, catalog ab1890), mouse anti–β-actin Ab (1:2,000, Abcam, catalog ab6276) or goat anti-GAPDH Ab (1:2,000, Abcam, catalog ab9483) was used.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Co-Immunoprecipitation Assay, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: MicroRNA-210 overexpression promotes psoriasis-like inflammation by inducing Th1 and Th17 cell differentiation

doi: 10.1172/JCI97426

Figure Lengend Snippet: In psoriasis patients, inflammatory factors TGF-β and IL-23 stimulate the expression of HIF-1α, which recruits P300 to the miR-210 promoter region and increases the H3ac levels on miR-210 promoters, thus leading to the upregulation of miR-210 expression. miR-210 represses STAT6 and LYN expression through binding the 3′-UTRs of STAT6 and LYN mRNA, which promotes Th1/Th17 cell differentiation and inhibits Th2 cell differentiation in psoriasis. Moreover, miR-210 overexpression also enhances the proliferation and chemokine secretion of keratinocytes, which increases the recruitment of activated T cells in skin lesions. Together, the effects of the increased miR-210 in CD4+ T cells and keratinocytes contribute to the formation of psoriatic skin lesions.

Article Snippet: Rabbit anti–HIF-1α Ab (1:1,000; Cell Signal Technology, catalog 14179), rabbit anti-STAT6 Ab (1:1,000; Cell Signal Technology, catalog 9362), mouse anti-LYN Ab (1:1,000; Abcam, catalog ab1890), mouse anti–β-actin Ab (1:2,000, Abcam, catalog ab6276) or goat anti-GAPDH Ab (1:2,000, Abcam, catalog ab9483) was used.

Techniques: Expressing, Binding Assay, Cell Differentiation, Over Expression