Journal: Journal of Biological Chemistry

Article Title: Tristetraprolin-dependent Post-transcriptional Regulation of Inflammatory Cytokine mRNA Expression by Apolipoprotein A-I

doi: 10.1074/jbc.m110.202275

Figure Lengend Snippet: FIGURE 4. STAT3 activation is involved in the apoA-I-mediated increase of TTP expression in LPS-treated macrophages. A, THP-1 macrophages were treated with apoA-I for different times as indicated. Proteins were extracted, and the phosphorylated STAT1, STAT3, and STAT6 levels were measured by immunoblot analyses with antibodies specific for phosphorylated signal transducer and activator of transcriptions. Values were normalized against -actin. *, p 0.05 versus 0 min. B, THP-1 macrophages were treated with apoA-I for different times as indicated. The nuclear proteins extracted from cells were subjected to immunoblot analyses with antibodies against STAT3 and histone H1. *, p 0.05 versus 0 min. C, THP-1 macrophages were pretreated with apoA-I for 30 min. Thereafter, the medium was replaced with fresh medium containing the AG-490 (30 M). Then cells were incubated for another 30 min, and total or nuclear proteins were subjected to immunoblot analyses with antibody against p-JAK2 and STAT3. *, p 0.05. D, THP-1 macrophages were transfected with control (WT) or STAT3 siRNA for 48 h, and protein samples were immunoblotted with STAT3 and -actin antibodies. *, p 0.05 versus control group. E, THP-1 macrophages transfected with control or STAT3 siRNA were incubated with LPS for 4 h with or without pretreatment of apoA-I. Protein samples were immunoblottedwithTTPand-actinantibodies.*,p 0.05versuscontrolgroup.F,THP-1macrophageswerepretreatedwithAG-490orDMSOfor1h,andcells were then incubated with LPS for another 4 h with or without pretreatment of apoA-I. Protein samples were immunoblotted with TTP and -actin antibodies. *, p 0.05 compared with DMSO group. All of the results are the mean S.D. of quadruplicate values from three separate experiments.

Article Snippet: Antibodies and Reagents—STAT1, phospho-STAT1, STAT6, phospho-STAT6, TTP, HuR, TNF- , and COX-2 antibodies and histone H1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Activation Assay, Expressing, Western Blot, Incubation, Transfection, Control

Journal: Molecules

Article Title: Piperine Ameliorates Trimellitic Anhydride-Induced Atopic Dermatitis-Like Symptoms by Suppressing Th2-Mediated Immune Responses via Inhibition of STAT6 Phosphorylation

doi: 10.3390/molecules25092186

Figure Lengend Snippet: Effect of piperine on TMA-induced Th2-associated immune responses in dLNs. The dLNs were seeded to 1 × 10 6 cells/mL and cultured in the presence of Con A (2 μg/mL) for 48 h. ( A ) The secretion of IL-4 cytokine, ( B ) the mRNA levels of GATA3, and ( C ) STAT6 phosphorylation were measured by ELISA, RT-PCR, and Western Blot assay, respectively. The results are shown as the means ± SD ( n = 3). Asterisks (*) and (**) indicate significant differences between piperine-treated groups and sham groups of TMA-induced AD-like mice at p < 0.05 and p < 0.01, respectively.

Article Snippet: Antibodies specific for STAT6, β-actin, horseradish peroxidase (HRP)-conjugated goat anti-rabbit, anti-mouse antibodies were purchased from Santa Cruz (CA, USA).

Techniques: Cell Culture, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Molecules

Article Title: Piperine Ameliorates Trimellitic Anhydride-Induced Atopic Dermatitis-Like Symptoms by Suppressing Th2-Mediated Immune Responses via Inhibition of STAT6 Phosphorylation

doi: 10.3390/molecules25092186

Figure Lengend Snippet: Effect of piperine treatment on IL-4 production in OVA-immunized splenocytes and on IL-4-mediated STAT6 phosphorylation in CD4 + T cells. ( A ) For the analysis of IL-4 cytokine production, the splenocytes isolated from OVA-immunized BALB/c mice were seeded to 5 × 10 6 cells/mL and cultured in the presence or absence of OVA (100 μg/mL) with piperine. ( B ) To detect STAT6 phosphorylation, CD4 + T cells isolated from splenocytes of a BALB/c mouse were pre-treated with piperine for 1 h before IL-4 (20 ng/mL) treatment for 15 min. The results are shown as the means ± SD ( n = 3). Asterisks (**) indicate significant differences between the piperine-treated and non-treated groups at p < 0.01.

Article Snippet: Antibodies specific for STAT6, β-actin, horseradish peroxidase (HRP)-conjugated goat anti-rabbit, anti-mouse antibodies were purchased from Santa Cruz (CA, USA).

Techniques: Phospho-proteomics, Isolation, Cell Culture

Journal: Molecules

Article Title: Piperine Ameliorates Trimellitic Anhydride-Induced Atopic Dermatitis-Like Symptoms by Suppressing Th2-Mediated Immune Responses via Inhibition of STAT6 Phosphorylation

doi: 10.3390/molecules25092186

Figure Lengend Snippet: Effects of piperine on TMA-induced infiltration of CCR3 + cells in mouse ear tissues and IL-4-induced CCL26 mRNA expression and STAT6 phosphorylation in HaCaT cells. HaCaT cells pre-incubated with piperine were stimulated with IL-4 for 15 min and then incubated for 24 h to analyze ( A ) CCL26 mRNA expression and ( B ) STAT6 phosphorylation relatively. ( C ) Infiltration of CCR3 + cells into ear tissue was observed by immunohistological analysis. Black arrows indicate CCR3 + cells. The results are shown as the means ± SD ( n = 3). Asterisks (**) indicate significant differences between the piperine-treated and non-treated groups at p < 0.01.

Article Snippet: Antibodies specific for STAT6, β-actin, horseradish peroxidase (HRP)-conjugated goat anti-rabbit, anti-mouse antibodies were purchased from Santa Cruz (CA, USA).

Techniques: Expressing, Phospho-proteomics, Incubation