Journal: PLoS ONE

Article Title: HDL Induces the Expression of the M2 Macrophage Markers Arginase 1 and Fizz-1 in a STAT6-Dependent Process

doi: 10.1371/journal.pone.0074676

Figure Lengend Snippet: BMDM were grown as previously described and stimulated for 15min with either HDL alone (50μg/mL), IL-4 alone (10ng/mL) or both. Phosphorylation state for STAT6 (A) and STAT3 (C) was assessed by western blot. As a positive control for STAT3 activation, cells were treated with IL-10 (10ng/mL) for 30 min. Signal quantification for phospho-STAT6 (B) is the result of three independent experiments. Asterisks indicate statistically significant difference, either compared to control, or between 2 conditions when linked by a bar (*p<0.05, **p<0.01).

Article Snippet: The following antibodies were used: Phospho-STAT3 (Tyr705) and total-STAT3 (Cell Signaling), Phospho-STAT6 (Tyr641) (Imgenex), STAT6 (Santa Cruz Biotechnology Inc.), and GAPDH (Millipore).

Techniques: Phospho-proteomics, Western Blot, Positive Control, Activation Assay, Control

Journal: PLoS ONE

Article Title: HDL Induces the Expression of the M2 Macrophage Markers Arginase 1 and Fizz-1 in a STAT6-Dependent Process

doi: 10.1371/journal.pone.0074676

Figure Lengend Snippet: BMDM were isolated from STAT6 +/+ (black bars) and STAT6-/- (hatched bars) mice and were stimulated for 6h with HDL (50μg/mL) or IL-4 (10ng/mL). The induction of Arg-1 or Fizz-1 mRNA in the STAT6+/+ cells by HDL was ~3X for each and by IL-4, ~1800 or 3300, respectively; these values were set to 100%. Results are representative of three independent experiments. Asterisks indicate statistically significant differences compared to corresponding STAT6 +/+ conditions (***p<0.001).

Article Snippet: The following antibodies were used: Phospho-STAT3 (Tyr705) and total-STAT3 (Cell Signaling), Phospho-STAT6 (Tyr641) (Imgenex), STAT6 (Santa Cruz Biotechnology Inc.), and GAPDH (Millipore).

Techniques: Isolation

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 1 STAT6 is activated in the kidneys of UUO and HFD mice. A Representative micrographs for IHC staining of p-STAT6 in kidney section and quantification are performed. B The mRNA and C protein expression of STAT6, p-STAT6, Arg-1, TGF-β, α-SMA, FN in the indicated groups were determined by qRT-PCR or immunoblot analyses with the quantification on the right panel. D Representative micrographs for Sirius red and relative collagen proportion was quantified. E Representative micrographs for H&E and Oil Red O staining in kidney sections from indicated group. F Kidney TG content were measured in the indicated groups. Results are expressed as the mean ± SD (n = 8, *p < 0.05, Ctrl vs. Treatments). G Nuclear proteins were extracted from kidneys from different individuals as indicated, STAT6 and H3 were assayed by western blot analysis.

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Western Blot, Staining

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 2 STAT6 deficiency in tubular cells attenuates UUO-caused renal fibrosis. The tubular-specific-Stat6 KO and control littermates were sacrificed 1w after UUO operation. A Representative IHC staining of p-STAT6, α-SMA, and micrographs for H&E, Sirius red staining in the control and fibrotic kidneys from the indicated group and quantification was performed. BThe mRNA and C protein expression of p-STAT6, STAT6, Arg-1, TGF-β, α-SMA, FN in the indicated treatments were determined by qRT-PCR or immunoblot analyses with the quantification on the right panel. Results are expressed as the mean ± SD (n = 8, *p < 0.05, Stat6 WT vs. Stat6 cKO).

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

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

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 3 STAT6 deficiency in tubular cells attenuates UUO-caused lipid accumulation. The tubular-specific-Stat6 KO and control littermates were sacrificed 1w after UUO operation. A Representative IHC staining of PPARα and Oil Red O staining from Stat6 WT and cKO kidneys with or without UUO, and quantification for PPARα was performed. B TG content were determined in the kidneys from Stat6 WT and cKO mice with or without UUO. C mRNA levels of genes related to lipid metabolism in Stat6 WT and cKO mice with or without UUO operation. Results are expressed as the mean ± SD (n = 8, *p < 0.05, Stat6 WT vs. Stat6 cKO).

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

Techniques: Control, Immunohistochemistry, Staining

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 4 STAT6 in tubular cells suppresses PPARα-mediated fatty acid oxidation. HK2 cells were transfected with siRNA or plasmid for STAT6 inhibition or overexpression. Followed by 24 h serum-free medium culture, the cells were treated with TGF-β (5 ng/ml) for another 24 h. A Representative traces of three independent experiments for the measurement of oxygen consumption rate (OCR) were shown. Oligomycin, FCCP, and antimycin/rotenone were administrated at the indicated time point. ATP production was determined according to the OCR values. Data are presented as mean ± SD (n = 3, *P < 0.05). B Representative micrographs for Oil Red O staining of HK2 cells treated as indicated (C) TG content was determined enzymatically. D, E mRNA levels of genes related to lipid metabolism in STAT6 inhibited or overexpressed HK2 cells were determined by qRT-PCR. Results are expressed as the mean ± SD (n = 4 *p < 0.05, SiCtrl/Vector vs. SiSTAT6/STAT6 transfection) F mRNA levels of genes related to FAO and fibrotic proteins expression in primary renal tubular epithelial cells isolated from the above Stat6 WT and cKO mice. Results are expressed as the mean ± SD (n = 4, *p < 0.05, Stat6 WT vs. Stat6 cKO). G HK2 cells were transfected with indicated siRNA or plasmid for 24 h in serum-free medium and followed by TGF-β (5 ng/ml) treatment for another 24 h. Cell lysates were harvested and subjected to immunoblot analyses with the indicated antibodies. Quantification of relative protein expression was determined. Results are expressed as the mean ± SD (n = 4, *p < 0.05, Ctrl vs. TGF-β; #p < 0.05, SiCtrl/Vector vs.SiSTAT6/STAT6 transfection).

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

Techniques: Transfection, Plasmid Preparation, Inhibition, Over Expression, Staining, Quantitative RT-PCR, Expressing, Isolation, Western Blot

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 5 STAT6 regulates FAO and fibrotic related protein expression by targeting PPARα in tubular cells. HK2 cells were transfected with siRNA or plasmid for STAT6 or PPARα. After 24 h incubation, cells were treated with TGF-β (5 ng/ml) for another 24 h. A Cell lysates were harvested and subjected to immunoblot analyses with the indicated antibodies. Quantification of relative protein expression was determined. Results are expressed as the mean ± SD (n = 4 *p < 0.05, SiCtrl/Vector vs. siSTAT6/STAT6 transfection; #p < 0.05, SiCtrl/Vector vs. SiPPARα/PPARα transfection). B Representative micrographs showing the Oil Red O staining of indicated treatments in HK2 cells. C Identification of four SIEs in the promoter of PPARα. The potential site of STAT6 binding to PPARα promoter with detected by ChIP assay in HK2 cells with or without TGF-β (5 ng/ml) 24 h treatment. Results are expressed as the mean ± SD (n = 4 *p < 0.05, IgG vs. STAT6 immunoprecipitation). D The different human PPARα promoter constructs were cloned upstream of a luciferase reporter gene. HK2 cells were either transfected with empty vector or these constructs along with Renilla luciferase reporter for 24 h and followed by another 24 h TGF-β treatment, dual-luciferase activities were measured. The experiment was repeated three times, each with triplicate samples. Data are expressed as mean ± SD (n = 3, *p < 0.05, Vector vs. PPARα promoter constructs). E, F HK2 cells were transfected with indicated siRNA and plasmid for 24 h in serum-free medium, followed by TGF-β (5 ng/ml) 24 h and harvested for qRT-PCR analysis of the indicated genes. Results are expressed as the mean ± SD (n = 4 *p < 0.05, SiCtrl/ Vector vs. siSTAT6/STAT6 transfection; #p < 0.05, SiCtrl/Vector vs. SiPPARα/PPARα transfection).

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

Techniques: Expressing, Transfection, Plasmid Preparation, Incubation, Western Blot, Staining, Binding Assay, Immunoprecipitation, Construct, Clone Assay, Luciferase, Quantitative RT-PCR

Journal: Cell death & disease

Article Title: STAT6 contributes to renal fibrosis by modulating PPARα-mediated tubular fatty acid oxidation.

doi: 10.1038/s41419-022-04515-3

Figure Lengend Snippet: Fig. 6 AS1517499 attenuates aberrant lipid metabolism and tubulointerstitial fibrosis in UUO mice. A Representative micrographs for p-STAT6 staining in the kidney from the indicated groups, and quantification of relative protein expression in the kidneys from the indicated groups. B Representative Sirius red staining and H&E staining in the kidney from the indicated groups, and quantification of relative collagen proportion in the kidneys from the indicated groups. C Representative Oil O Red staining in the kidney from the indicated groups. D TG content was determined in the kidneys from the indicated groups. E Kidney tissue lysates from each group were subjected to immunoblot analyses with the indicated antibodies. Representative blots of three independent samples in each group were shown and quantification of relative protein expression was determined. F The mRNA levels of genes related to lipid metabolism and renal fibrosis in the kidneys from the indicated groups. Results are expressed as the mean ± SD (n = 6-8, *p < 0.05, UUO vs. Ctrl, #p < 0.05, UUO vs. UUO + AS1517499).

Article Snippet: The following antibodies were used: STAT6 (sc-374021), p-STAT6 (sc-136019), Arg-1 (sc166920), TGF-β (sc-146), α-SMA (sc-53142), FN (sc-18827), H3 (sc-517576), GAPDH (sc-32233) from Santa Cruz Biotechnology.

Techniques: Staining, Expressing, Western Blot

Journal: Cell reports

Article Title: Mass Cytometry Reveals Global Immune Remodeling with Multi-lineage Hypersensitivity to Type I Interferon in Down Syndrome

doi: 10.1016/j.celrep.2019.10.038

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-human phospho-STAT6 (Tyr641) (clone 18) , Fluidigm , Cat#3168012A; RRID:AB_2811103.

Techniques: Recombinant, Mass Cytometry, Software