Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Primers for qRT-PCR.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques:

Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Effect of SPI on JAK1-STAT6 signaling and TGFβ levels in mouse ileum tissue. ( A ) qRT-PCR analysis of Jak1 , Stat6 , Tgfb1 , and Tgfb2 mRNA expression levels in mouse ileum tissue (normalized against Actb (β-actin); n = 8). ( B ) Western blotting analysis of JAK1, p -JAK1, STAT6, p -STAT6, pIgR, and TGFβ1 protein levels in mouse ileum tissue ( n = 6). ( C ) The statistical analyses of the Western blotting results. Results are normalized to β-actin ( n = 6). All data are presented as means ± SEM. NS, not significant ( p > 0.05); * p < 0.05; ** p < 0.01. Statistical significance was calculated using t -tests.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Journal of Neurotrauma

Article Title: Microglial/Macrophage Polarization Dynamics following Traumatic Brain Injury

doi: 10.1089/neu.2015.4268

Figure Lengend Snippet: M1-like and mixed transitional (Mtran) microglial/macrophages predominate the peri-lesional cortex at 7 days post-injury. Representative images and analysis of M1- and M2-like polarized microglia/macrophages after controlled cortical impact (CCI). (A) Transforming growth factor (TGF) β+ (red) and CD16/32+ (green) microglia/macrophages in the peri-lesional and distal cortex at 24 h and 7 days post-injury. Insets display M1-like (CD16/32+), M2-like (TGFβ+), and Mtran (CD16/32+/TGFβ+) cells at each time-point. Scale bar = 50 μm. (B) Quantification of CD16/32+, TGFβ+, and CD16/32+/TGFβ+ cells at 24 h and 7 days post-injury. (C) Arginase 1+ (magenta) and inducible nitric oxide synthase (iNOS)+ (green) microglia/macrophages in the peri-lesional and distal cortex at 24 h and 7 days post-injury. Insets display M1-like (iNOS+) and Mtran (iNOS+/Arg1+) cells at 7 days post-injury. Scale bar = 50 μm (D) Quantification of iNOS+, Arg1+, and iNOS+/Arg1+ cells at 24 h and 7 days post-injury. Protein expression levels determined by binary area per region of interest (ROI) (mm2). n = 4 per time-point. Scale bar = 50 μm. Color image is available online at www.liebertpub.com/neu

Article Snippet: Briefly, 20-μm brain sections were washed three times with 1× PBS, blocked for 1 h in goat serum containing 0.4% Triton X-100, and incubated overnight at 4°C with a combination of primary antibodies, including rat anti-CD16/32 (1:1000, BD Biosciences), rabbit anti-TGFβ (1:500; Torrey Pines Biolabs Inc., Secaucus, NJ), rabbit anti-iNOS (1:400; Calbiochem, San Diego, CA), goat anti-arginase 1 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-gp91 phox (1:200; BD Transduction Laboratories), rat anti-CD68 (1:200; AbD Serotec, Raleigh, NC), rabbit anti-Iba1 (1:200; Wako Chemicals, Richmond, VA), rabbit anti-H2AX (1:2000; Novus Biologicals, Littleton, CO).

Techniques: Expressing

Journal: Journal of Neurotrauma

Article Title: Microglial/Macrophage Polarization Dynamics following Traumatic Brain Injury

doi: 10.1089/neu.2015.4268

Figure Lengend Snippet: NADPH oxidase (NOX2) co-localizes with M1-like and mixed transitional (Mtran) microglia/macrophages at 7 days post-injury. Representative images of NOX2 expression with M1-like, M2-like, and Mtran microglia/macrophages after controlled cortical impact (CCI). (A) gp91phox+ (red), CD16/32+ (magenta), and transforming growth factor (TGF) β+ (green) cells in the peri-lesional cortex (upper panels) and at distant subcortical sites (lower panel). Insets display co-localization of gp91phox with CD16/32+ and CD16/32+/TGFβ+ cells in peri-lesional regions, and lack of gp91phox expression in TGFβ+ only cells in distal regions. (B) gp91phox+ (red), iNOS+ (green), and Arg1+ (magenta) cells in the peri-lesional cortex (upper panels) and at distant subcortical sites (lower panel). Insets display co-localization of gp91phox with inducible nitric oxide synthase (iNOS)+ and iNOS+/Arg1+ cells in peri-lesional regions, and lack of gp91phox expression in Arg1+ only cells in distal regions. Scale bar = 50 μm. (C) Quantification of co-localization of gp91phox+ with CD16/32+, TGFβ+, and CD16/32+/TGFβ+ in the peri-lesional and distal cortex at 24 h and 7 days post-injury. (D) Quantification of co-localization of gp91phox+ with iNOS+, Arg1+, and iNOS+/Arg1+ in the peri-lesional and distal cortex at 24 h and 7 days post-injury. Protein expression levels determined by binary area per region of interest (ROI; mm2). n = 4 per time-point. Color image is available online at www.liebertpub.com/neu

Article Snippet: Briefly, 20-μm brain sections were washed three times with 1× PBS, blocked for 1 h in goat serum containing 0.4% Triton X-100, and incubated overnight at 4°C with a combination of primary antibodies, including rat anti-CD16/32 (1:1000, BD Biosciences), rabbit anti-TGFβ (1:500; Torrey Pines Biolabs Inc., Secaucus, NJ), rabbit anti-iNOS (1:400; Calbiochem, San Diego, CA), goat anti-arginase 1 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-gp91 phox (1:200; BD Transduction Laboratories), rat anti-CD68 (1:200; AbD Serotec, Raleigh, NC), rabbit anti-Iba1 (1:200; Wako Chemicals, Richmond, VA), rabbit anti-H2AX (1:2000; Novus Biologicals, Littleton, CO).

Techniques: Expressing

Journal: Journal of Neurotrauma

Article Title: Microglial/Macrophage Polarization Dynamics following Traumatic Brain Injury

doi: 10.1089/neu.2015.4268

Figure Lengend Snippet: Dinucleotide phosphate oxidase (NOX2) inhibition alters microglial/macrophage polarization after controlled cortical impact (CCI). (A) Experimental design for gp91ds-tat intervention study. (B) Image analysis was performed in two regions of interest (ROIs) that included peri-lesional cortex (ROI1) and the surrounding cortex distal to the lesion (ROI2). (C) Representative images of gp91phox and CD68 expression in ds-tat scrambled peptide control-treated and gp91ds-tat-treated CCI mice at 7 days post-injury. gp91phox+ (green) and CD68+ (red) cells in ROI1 and ROI2. Scale bar = 50 μm. (D) Representative images of CD16/32 and transforming growth factor (TGF) β expression in ds-tat scrambled peptide control-treated and gp91ds-tat-treated CCI mice at 7 days post-injury. CD16/32+ (magenta) and TGFβ+ (green) cells in the ipsilateral cortex. Scale bar = 50 μm. (E) Quantification of expression of gp91phox and CD68 in ROI1 and ROI2 of the ipsilateral cortex at 7 days post-injury. (F) Quantification of expression of CD16/32+, TGFβ+, and CD16/32+/TGFβ+ in the peri-lesional cortex at 7 days post-injury. Protein expression levels determined by binary area per ROI (mm2). n = 4 per time-point. Color image is available online at www.liebertpub.com/neu

Article Snippet: Briefly, 20-μm brain sections were washed three times with 1× PBS, blocked for 1 h in goat serum containing 0.4% Triton X-100, and incubated overnight at 4°C with a combination of primary antibodies, including rat anti-CD16/32 (1:1000, BD Biosciences), rabbit anti-TGFβ (1:500; Torrey Pines Biolabs Inc., Secaucus, NJ), rabbit anti-iNOS (1:400; Calbiochem, San Diego, CA), goat anti-arginase 1 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-gp91 phox (1:200; BD Transduction Laboratories), rat anti-CD68 (1:200; AbD Serotec, Raleigh, NC), rabbit anti-Iba1 (1:200; Wako Chemicals, Richmond, VA), rabbit anti-H2AX (1:2000; Novus Biologicals, Littleton, CO).

Techniques: Inhibition, Expressing

Journal: Naunyn-Schmiedeberg's archives of pharmacology

Article Title: Alleviation of pulmonary fibrosis by the dual PPAR agonist saroglitazar and breast milk mesenchymal stem cells via modulating TGFß/SMAD pathway.

doi: 10.1007/s00210-024-03004-y

Figure Lengend Snippet: Fig. 3 Effect of BrMSCs and/or saroglitazar on lipid peroxidation parameters A MDA and B SOD, the pro-inflammatory cytokines C IL-1β, D IL-10, E TGF-ß, and F serum Beclin-1 in rats with BLM-induced lung fibrosis

Article Snippet: To localize alpha-smooth muscle actin (α-SMA) protein as a marker for myofibroblasts, anti-alpha-smooth muscle actin antibody (rabbit polyclonal antibody; No. ab5694; dilution 1/50; Abcam, Cambridge, UK) (Egger et al. 2013) and with a monoclonal rabbit anti-TGFβ1 antibody (Clone TB21, MCA797, Serotec, Oxford, UK) at 1:200 dilution for TGF-β1 detection (Attia et al. 2017).

Techniques:

Journal: Naunyn-Schmiedeberg's archives of pharmacology

Article Title: Alleviation of pulmonary fibrosis by the dual PPAR agonist saroglitazar and breast milk mesenchymal stem cells via modulating TGFß/SMAD pathway.

doi: 10.1007/s00210-024-03004-y

Figure Lengend Snippet: Fig. 4 Effect of BrMSCs and/or saroglitazar on the mRNA expression genes of A TGF-β, B SMAD-3, C SMAD-7, D PPAR-γ, E PPAR-α in rats’ lungs with BLM-induced pulmonary fibrosis

Article Snippet: To localize alpha-smooth muscle actin (α-SMA) protein as a marker for myofibroblasts, anti-alpha-smooth muscle actin antibody (rabbit polyclonal antibody; No. ab5694; dilution 1/50; Abcam, Cambridge, UK) (Egger et al. 2013) and with a monoclonal rabbit anti-TGFβ1 antibody (Clone TB21, MCA797, Serotec, Oxford, UK) at 1:200 dilution for TGF-β1 detection (Attia et al. 2017).

Techniques: Expressing

Journal: Naunyn-Schmiedeberg's archives of pharmacology

Article Title: Alleviation of pulmonary fibrosis by the dual PPAR agonist saroglitazar and breast milk mesenchymal stem cells via modulating TGFß/SMAD pathway.

doi: 10.1007/s00210-024-03004-y

Figure Lengend Snippet: Fig. 10 A photomicrograph of immune-stained sections of the study groups. A CN-G showing faint expression of TGF-β1 by some cells in the alveolar septa between the alveoli and alveolar sacs. B, C BLM-G sections showing [B] Numerous cytoplasmic reactions for TGF- β1 positive immune expressed cells (arrows) within the inter- alveolar septum. C Increased TGF-β1 positive immune expressed cells (arrows) within the inter-alveolar septum. D BrMSCs-G showing some positive immune expressed cells (arrows) for TGF-β1 within the inter-alveolar septum. E Saro-G showing few positive immune expressed cells (arrows) for TGF-β1 within the inter-alveo- lar septum. F BrMSCs + Saro-G showing faint weak positive TGF-β1 immune expressed cells (arrows) within the inter- alveolar septum. (TGF-β1 X 400) (scale bar = 40 μm). G Mean values of % area of TGF-ß1 among different studied groups, one way ANOVA was used for statistical analysis. “a” means value is statistical significance when comparing CN-G with other groups, “b” means the value is statistical significance when comparing BLM-G with other groups, “c” means the value is statistical significance when comparing BrMSCs + Saro-G with other groups. The same letters mean no statistical significance dif- ference

Article Snippet: To localize alpha-smooth muscle actin (α-SMA) protein as a marker for myofibroblasts, anti-alpha-smooth muscle actin antibody (rabbit polyclonal antibody; No. ab5694; dilution 1/50; Abcam, Cambridge, UK) (Egger et al. 2013) and with a monoclonal rabbit anti-TGFβ1 antibody (Clone TB21, MCA797, Serotec, Oxford, UK) at 1:200 dilution for TGF-β1 detection (Attia et al. 2017).

Techniques: Staining, Expressing

Journal: Journal of cell science

Article Title: Integrin alpha8beta1 mediates adhesion to LAP-TGFbeta1.

doi: 10.1242/jcs.00145

Figure Lengend Snippet: Fig. 2. (i) α8β1 mediates spreading on LAP-TGFβ1. α8-transfected or mock-transfected AtT20 cells were plated on 5 µg/ml of recombinant LAP-TGFβ1 in serum-free media for 24 or 48 hours. At 24 hours, mock transfected AtT20 cells remain rounded and unattached on LAP-TGFβ1 (A), whereas α8-transfected cells attach and spread on LAP-TGFβ1 (B). Further spreading is seen at 48 hours (C). (ii) Adhesion of α8β1 to LAP-TGFβ1 results in FAK phosphorylation. α8-transfected or mock-transfected AtT20 cells were plated on 0.01% poly-L-lysine (PLL), 5 µg/ml fibronectin (FN), 5 µg/ml LAP-TGFβ1 (LAP1) or 5 µg/ml RGE-LAP-TGFβ1 (RGE) for 30 minutes in serum-free media. Cells were lysed in buffer containing phosphatase inhibitors, immunoprecipitated with anti- FAK antibody followed by western blotting with either anti- phosphotyrosine antibody PY20 (top panel) or anti-FAK antibody (bottom panel).

Article Snippet: Wells were washed with PBS0.05% Tween and then incubated with 50 μl of anti-LAP-TGFβ1 IgG antibody (0.5 μg/ml) (R&D Systems) at RT for 2 hours.

Techniques: Transfection, Recombinant, Phospho-proteomics, Immunoprecipitation, Western Blot

Journal: Journal of cell science

Article Title: Integrin alpha8beta1 mediates adhesion to LAP-TGFbeta1.

doi: 10.1242/jcs.00145

Figure Lengend Snippet: Fig. 3. (A) Proliferation of cells adherent to LAP-TGFβ1. Equal numbers of α8-transfected or mock-transfected AtT20 cells were plated on 5 µg/ml LAP-TGFβ1 or 5 µg/ml fibronectin (FN) in serum-free media. After 3 days, proliferation was assayed using the Roche Cell Proliferation kit (MTT). Results for four independent clones of AtΤ20α8 cells are shown. Data is reported as the mean absorbance of triplicate wells±s.d. (B) ERK phosphorylation on LAP-TGFβ1. α8-transfected or mock-transfected AtT20 cells were plated on 0.01% poly-L-lysine (PLL) 5 µg/ml fibronectin (FN), 5 µg/ml LAP-TGFβ1 (LAP1), or 5 µg/ml RGE-LAP-TGFβ1 (RGE) for 30 minutes in serum-free media and then lysed in buffer containing phosphatase inhibitors. Equal amounts of protein were loaded and probed with an antibody to phospho-ERK (top) or ERK (bottom).

Article Snippet: Wells were washed with PBS0.05% Tween and then incubated with 50 μl of anti-LAP-TGFβ1 IgG antibody (0.5 μg/ml) (R&D Systems) at RT for 2 hours.

Techniques: Transfection, Clone Assay, Phospho-proteomics

Journal: Journal of cell science

Article Title: Integrin alpha8beta1 mediates adhesion to LAP-TGFbeta1.

doi: 10.1242/jcs.00145

Figure Lengend Snippet: Fig. 5. LAP-TGFβ1 localization in adult lung tissue. Paraffin- embedded, formalin fixed tissues were stained with an antibody specific for LAP-TGFβ1 and counterstained with hematoxylin. Immunoreactivity for LAP-TGFβ1 is seen on a macrophage (arrowhead) and interstitial cells (arrows). Magnification: A, 4×; B,C, 20×; D, 40×.

Article Snippet: Wells were washed with PBS0.05% Tween and then incubated with 50 μl of anti-LAP-TGFβ1 IgG antibody (0.5 μg/ml) (R&D Systems) at RT for 2 hours.

Techniques: Staining

Journal: Renal Failure

Article Title: Increased expression of the P2Y ₁₂ receptor is involved in the failure of autogenous arteriovenous fistula caused by stenosis

doi: 10.1080/0886022x.2023.2278314

Figure Lengend Snippet: Figure 1. Hematoxylin-eosin (HE) and Elastic-Van Gieson (EVG) staining, and immunohistochemical staining for P2Y12, TGF-β1, MCP-1, and CD68 in the venous tissues between the two groups. (A) Comparison of HE and EVG staining results in venous tissues between the two groups. Scale bar: 200 μm. (B and C) Immunohistochemical staining for P2Y12, TGF-β1, MCP-1, and CD68 in the venous tissues of both groups. Representative images of P2Y12, TGF-β1, MCP-1, and CD68 staining are presented (B). semiquantitative analysis results revealed a significant increase in the expression of P2Y12, TGF-β1, MCP-1, and CD68 in the experimental group as compared to that in the control group (C). two-sample t-tests were performed for statistical analysis (C). Each bar represents mean ± standard error of the mean (SEM). significant differences are denoted as **p < 0.01 and ****p < 0.0001. Scale bar: 100 μm. CG and EG refer to the control and experimental groups, respectively.

Article Snippet: Anti-P2Y12 rabbit polyclonal antibody (NBP1-78249) and anti-TGFβ1 rabbit polyclonal antibody (NBP1-80289) were purchased from Novus Bio (USA).

Techniques: Staining, Immunohistochemical staining, Comparison, Expressing, Control

Journal: Renal Failure

Article Title: Increased expression of the P2Y ₁₂ receptor is involved in the failure of autogenous arteriovenous fistula caused by stenosis

doi: 10.1080/0886022x.2023.2278314

Figure Lengend Snippet: Figure 2. The expression of P2Y12, TGF-β1, and MCP-1 was assessed by western blot and qRT-PCR. (A and B) The results showed that the expression of P2Y12, TGF-β1, and MCP-1 was significantly increased in stenotic venous tissues of the experimental group as compared to that in venous tissues of the control group. The quantitation of the expression ratio was determined by western blot (shown with representative images) and quantified via densitom etry analysis. (C) qRT-PCR analysis of P2Y12, TGF-β1, and MCP-1 expression in venous tissues of the control and experimental groups. The results showed that a significant increase in the expression of P2Y12, TGF-β1, and MCP-1 in stenotic venous tissues of the experimental group as compared to that in venous tissues of the control group. Two-sample t-test were performed for statistical analysis (B and C). Each bar represents mean ± SEM. Significant differ ences are denoted as *p < 0.05, **p < 0.01, and ****p < 0.0001. CG and EG refer to the control and experimental groups respectively.

Article Snippet: Anti-P2Y12 rabbit polyclonal antibody (NBP1-78249) and anti-TGFβ1 rabbit polyclonal antibody (NBP1-80289) were purchased from Novus Bio (USA).

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

Journal: Renal Failure

Article Title: Increased expression of the P2Y ₁₂ receptor is involved in the failure of autogenous arteriovenous fistula caused by stenosis

doi: 10.1080/0886022x.2023.2278314

Figure Lengend Snippet: Figure 5. Double immunofluorescent staining for the CD68 with α-SMA or P2Y12 with TGF-β1 in venous tissues between the two groups. (A) Representative photomicrographs showing CD68(red) and α-SMA (green) in venous tissues of the control group. White arrows point to CD68+/α-SMA+ cells. Scale bar: 50 μm. (B) Representative photomicrographs showing CD68(red) and α-SMA (green) in stenotic venous tissues of the experimental group. White arrows point to CD68+/α-SMA+ cells. Scale bar: 50 μm. (C) Representative photomicrographs showing P2Y12(red) and TGF-β1(green) in venous tissues of the control group. White arrows point to P2Y12

Article Snippet: Anti-P2Y12 rabbit polyclonal antibody (NBP1-78249) and anti-TGFβ1 rabbit polyclonal antibody (NBP1-80289) were purchased from Novus Bio (USA).

Techniques: Staining, Control