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 Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: The prognosis of gastric adenocarcinoma suggests that TGFβ1 has great potential. a The pan cancer expression of TGFβ1 in the TCGA database. b The pan cancer expression of TGFβ1 in paired samples in the TCGA database. c The expression of TGFβ1 in the GSE65801 dataset. d The expression of TGFβ1 in the GSE84787 dataset. e T staging results of TGFβ1. f The differential expression of TGFβ1 in the quality of life of different patients. g Comparing the prognosis between high and low TGFβ1 groups based on the g KM survival curve. h Analyze the predictive accuracy and efficacy of TGFβ1 in GC queue diagnosis. i Diagnostic prediction of TGFβ1 in different T stages of GC queue. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: Expressing, Quantitative Proteomics, Biomarker Discovery, Diagnostic Assay

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: The association between TGFβ1 and the immune microenvironment associated with GC tumors. a TGFβ1 pan cancer status and tumor associated immune cell correlation heatmap in TCGA database. b The expression of cell markers in the GSE65801 dataset. c GC tumor associated immune cells and TGFβ1 correlation bar chart. d Box plot of the correlation between TGFβ1 differential expression group and macrophages. e Scatter plot showing the association between TGFβ1 and macrophages. f The correlation results between TGFβ1 and GC immune infiltrating cells in TIMER database. g Correlation heatmap between TGFβ1 and macrophage subtypes. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: Expressing, Quantitative Proteomics

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: There is a close correlation between TGFβ1 and polarization of M2c macrophages. a The expression results of different biomarkers in the GSE21328 dataset. b The expression results of different biomarkers in the GSE70394 dataset. c The expression results of different biomarkers in the GSE103236 dataset. d Heat map of the association between TGFβ1 and different subtypes of macrophages. e The relationship between TGFβ1 and different subtypes of macrophage marker networks. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: Expressing, Marker

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: Different inducers promote polarization of different subtypes of macrophages. a The CCK-8 method was used to detect the survival of M0 macrophages after 24 h of IL-4 intervention. b The CCK-8 method was used to detect the survival of M0 macrophages after 24 h of intervention with TGFβ1. c The CCK-8 method was used to detect the survival of M0 macrophages after LPS intervention for 24 h. d RT-qPCR was used to detect the expression of CD86 mRNA. e RT-qPCR was used to detect the expression of iNOS mRNA. f RT-qPCR was used to detect the expression of CD206 mRNA. g RT-qPCR was used to detect the expression of IL1R2 mRNA. h RT-qPCR was used to detect the expression of CD163 mRNA. i RT-qPCR was used to detect the expression of TGFβ mRNA. j ELISA detects TNF - α levels. k ELISA was used to detect IL-10 levels. l ELISA detects the content of TGFβ1. m The WB results of different interventions on histone expression. f TNF-α protein expression results. f IL-10 protein expression results. f TGFβ1 protein expression results. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: CCK-8 Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: The effect of M2 subtype macrophages on TGFβ1 related pathway and epithelial mesenchymal transition in gastric cancer cells. a Cell intervention pattern diagram. b RT-qPCR was used to detect the levels of TGFβ mRNA in different intervention groups. c ELISA was used to test the expression of TNF-α in different intervention groups. d ELISA test the expression of IL-10 in different intervention groups. e ELISA was used to test the expression of TGFβ1 in different intervention groups. f The expression results of TGFβ1 protein. g The WB results of different interventions on histone expression. h The expression results of p-Smad2 protein. i The expression results of Smad2 protein. j The expression results of p-Smad3 protein. k The expression results of Smad3 protein. l The expression results of E-cadherin protein. m Results of N-cadherin protein expression. n Results of Vimentin protein expression. o RT-qPCR was used to detect the content of TGFβ mRNA at different intervention times. p ELISA was used to test the expression of TNF-α at different intervention times. q ELISA test the expression of IL-10 at different intervention times. r ELISA was used to test the expression of TGFβ1 at different intervention times. s WB results of protein expression at different intervention times. t The expression results of E-cadherin protein. u The expression results of N-cadherin protein. v Results of Vimentin protein expression. w Fluorescence results of mitochondrial membrane potential at different intervention times. Scale bar=50 μm. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Fluorescence, Membrane

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: M2c macrophages increase ferroptosis resistance in gastric cancer cells. a The CCK-8 method was used to detect the survival of gastric cancer cells (Hgc27 and MKN45) intervened with RSL3 for 24 h. b The CCK-8 method was used to detect the survival of gastric cancer cells (Hgc27 and MKN45) intervened with Fer-1 for 24 h. c The expression of SOD in different intervention groups. d The expression of MDA in different intervention groups. e The expression of GSH in different intervention groups. f The expression of TGFβ1 protein WB in different cell lines. g The expression results of TGFβ1 protein. h The expression of key ferroptosis proteins WB in different cell lines. i The expression results of FSP1 protein. j Expression results of DHODH protein. k Expression results of GPX4 protein. l SLC7A11 protein expression results. m The intervention of RSL3 on the expression of key ferroptosis protein WB in different co culture groups. n The expression results of GPX4 protein. o SLC7A11 protein expression results. p The WB expression of key proteins involved in ferroptosis in different intervention groups. q The expression results of GPX4 protein. r The expression results of SLC7A11 protein. s Fluorescence results of mitochondrial membrane potential in different intervention groups. Scale bar=50 μm. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: CCK-8 Assay, Expressing, Co-Culture Assay, Fluorescence, Membrane

Journal: Journal of Cancer

Article Title: The mutualistic relationship between M2c macrophages of TGFβ1 induction and gastric cancer cells: the correlation between protective mechanisms in the tumor microenvironment and polarization of subtypes of cells

doi: 10.7150/jca.97784

Figure Lengend Snippet: The role of gastric cancer cells in transforming macrophages in the TME. a The expression of M1 macrophage marker proteins in different groups. b The expression of M2 macrophage marker proteins in different groups. c Differential gene expression between M0 and gastric cancer cell metabolite intervention group. d Differential gene expression between M2 and gastric cancer cell metabolite intervention group. e Intersection statistics of differentially expressed genes between M2 and gastric cancer cell metabolite intervention group. f Intersection gene enrichment statistics. g RT-qPCR was used to detect the expression of CD86 mRNA. h RT-qPCR was used to detect the expression of iNOS mRNA. i RT-qPCR was used to detect the expression of CD206 mRNA. j RT-qPCR was used to detect the expression of IL1R2 mRNA. k RT-qPCR was used to detect the expression of CD163 mRNA. l RT-qPCR was used to detect the expression of TGFβ mRNA. m ELISA was used to detect TNF-α levels. n ELISA detects IL-10 levels. o ELISA detects the content of TGFβ1. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: After 2 h of closure with skimmed milk in TBST, the membranes were analyzed using TNF-α (Boster, A00002-5), IL-10 (Boster, A00021-2), TGFβ1 (Boster, BM4876), Smad2 (Immunoway, YP1185), p-Smad2 (Bioss, bs-20341R), Smad3 (Immunoway, YT4334), p-Smad3 (Bioss, bs-5235R), E-cadherin (Immunoway, YM0207), N-cadherin (Immunoway, YM4937), Vimentin (Immunoway, YT4880), GAPDH (Boster, BA2913), GPX4 (Boster, BM5231), FSP1 (Proteintech, #20886-1-AP), DHODH (Boster, M04035-1) and SLC7A11 (Boster, BM5318) were incubated overnight at 4°C.

Techniques: Expressing, Marker, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay