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Cell Signaling Technology Inc shp 2 antibody

HDCA modulates Treg migration and atherosclerotic plaque composition via FXR signaling. ApoE−/− mice (C57BL/6J background, male, 8 weeks old) were fed a high-fat diet for 28 days to induce AS and subsequently received adoptive transfer of control or FXR-knockout (FXR KO) Treg cells generated by CRISPR/Cas9-mediated lentiviral transduction, with HDCA (30 μM) or vehicle treatment as indicated. (A) Representative Western blot analysis of FXR, PD-1, <t>SHP-2,</t> p-Raptor, RAC and IL-10R expression in isolated Treg cells from each group. (B) Oil Red O staining was performed to assess lipid accumulation in the aorta. (C) Masson's trichrome staining of aortic sections from FXR KO mice reveals comparable plaque area and collagen deposition in both HDCA-treated and untreated groups (magnification, 5 × ; scale bar, 1 mm). (D) Representative H&E images of aortic sections show the difference between untreated and HDCA-treated mice in the FXR KO groups (magnification, 5 × ; scale bar, 500 μm). Relative bar graphs show quantification of lesion area and lesion/media area ratio. (E) Confocal immunofluorescence was used to evaluate Foxp3+ Treg infiltration within atherosclerotic plaques (scale bar, 25 μm). (F) Immunohistochemistry images show Treg accumulation in the plaque area across all groups (magnification, 40 × ; scale bar, 100 μm). (G) Flow cytometry analysis of Treg proportions in aortic plaques and spleen from control and FXR KO mice, with or without HDCA treatment. (H) Western blot analysis of matrix remodeling-related proteins, including calpain 1 and matrix metalloproteinase 2, and the anti-inflammatory factor IL-10. Data are presented as mean ± SD (n = 5 biological replicates). Data with four groups were analyzed by one-way ANOVA with Tukey's post hoc test. Comparisons between two groups were performed using the non-parametric Mann-Whitney U test. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Shp 2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 349 article reviews

shp 2 antibody - by Bioz Stars, 2026-09

95/100 stars

Images

1) Product Images from "Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis"

Article Title: Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis

Journal: Redox Biology

doi: 10.1016/j.redox.2026.104096

Figure Legend Snippet: HDCA modulates Treg migration and atherosclerotic plaque composition via FXR signaling. ApoE−/− mice (C57BL/6J background, male, 8 weeks old) were fed a high-fat diet for 28 days to induce AS and subsequently received adoptive transfer of control or FXR-knockout (FXR KO) Treg cells generated by CRISPR/Cas9-mediated lentiviral transduction, with HDCA (30 μM) or vehicle treatment as indicated. (A) Representative Western blot analysis of FXR, PD-1, SHP-2, p-Raptor, RAC and IL-10R expression in isolated Treg cells from each group. (B) Oil Red O staining was performed to assess lipid accumulation in the aorta. (C) Masson's trichrome staining of aortic sections from FXR KO mice reveals comparable plaque area and collagen deposition in both HDCA-treated and untreated groups (magnification, 5 × ; scale bar, 1 mm). (D) Representative H&E images of aortic sections show the difference between untreated and HDCA-treated mice in the FXR KO groups (magnification, 5 × ; scale bar, 500 μm). Relative bar graphs show quantification of lesion area and lesion/media area ratio. (E) Confocal immunofluorescence was used to evaluate Foxp3+ Treg infiltration within atherosclerotic plaques (scale bar, 25 μm). (F) Immunohistochemistry images show Treg accumulation in the plaque area across all groups (magnification, 40 × ; scale bar, 100 μm). (G) Flow cytometry analysis of Treg proportions in aortic plaques and spleen from control and FXR KO mice, with or without HDCA treatment. (H) Western blot analysis of matrix remodeling-related proteins, including calpain 1 and matrix metalloproteinase 2, and the anti-inflammatory factor IL-10. Data are presented as mean ± SD (n = 5 biological replicates). Data with four groups were analyzed by one-way ANOVA with Tukey's post hoc test. Comparisons between two groups were performed using the non-parametric Mann-Whitney U test. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Techniques Used: Migration, Adoptive Transfer Assay, Control, Knock-Out, Generated, CRISPR, Transduction, Western Blot, Expressing, Isolation, Staining, Immunofluorescence, Immunohistochemistry, Flow Cytometry, MANN-WHITNEY

Incubation:

Article Title: Bispecific BCMA/CD24 CAR-T cells control multiple myeloma growth.
Article Snippet: .. Primary antibodies SHP-1 (Cell Signaling Technology, catalog # 3759, clone # C14H6, 1:1000), SHP−2 (Cell Signaling Technology, catalog # 3397, clone # D50F2, 1:1000), Phospho-SHP-1 (Tyr564) (Cell Signaling Technology, catalog # 8849, clone # D11G5, 1:1000), Phospho-SHP-2 (Tyr580) (Cell Signaling Technology, catalog # 5431, clone # D66F10, 1:1000), β-Actin (Cell Signaling Technology, catalog # 4967, 1:1000), were incubated overnight at 4 °C. .. Anti-rabbit IgG, HRP-linked antibody (Cell Signaling Technology, catalog # 7074, 1:1000) were incubated for 2 h. ECL Substrates (Bio-Rad, catalog # 1705060) were used for exposure.

Article Title: Bispecific BCMA/CD24 CAR-T cells control multiple myeloma growth
Article Snippet: .. Primary antibodies SHP-1 (Cell Signaling Technology, catalog # 3759, clone # C14H6, 1:1000), SHP−2 (Cell Signaling Technology, catalog # 3397, clone # D50F2, 1:1000), Phospho-SHP-1 (Tyr564) (Cell Signaling Technology, catalog # 8849, clone # D11G5, 1:1000), Phospho-SHP-2 (Tyr580) (Cell Signaling Technology, catalog # 5431, clone # D66F10, 1:1000), β-Actin (Cell Signaling Technology, catalog # 4967, 1:1000), were incubated overnight at 4 °C. .. Anti-rabbit IgG, HRP-linked antibody (Cell Signaling Technology, catalog # 7074, 1:1000) were incubated for 2 h. ECL Substrates (Bio-Rad, catalog # 1705060) were used for exposure.

Article Title: Chitinase 3-like-1 (CHI3L1) Inhibits Innate Anti-Tumor and Tissue Remodeling Immune Responses by Regulating CD47-SIRPα and CD24-Siglec10-Mediated Phagocytosis
Article Snippet: .. After blocking, the membranes were incubated with the primary antibodies against CD47 (Invitrogen, miap301), SIRPα (Invitrogen, PAS-97176), CD24 (eBioscience, M1/69), Siglec10 (R&D Systems, MAB7775), HMGB1 (Abcam, ab-79823), calreticulin (Invitrogen, 1G6A7); SHP-1 (Cell Signaling Technology, C14H6), and SHP-2 (Cell Signaling Technology, #3752) overnight at 4°C in TBST and 5% BSA. ..

Western Blot:

Article Title: SHP-1 Regulates CD8+ T Cell Effector Function but Plays a Subtle Role with SHP-2 in T Cell Exhaustion Due to a Stage-Specific Nonredundant Functional Relay.
Article Snippet: RESEARCH ARTICLE | DECEMBER 13 2023 SHP-1 Regulates CD8+ T Cell E ector Function but Plays a Subtle Role with SHP-2 in T Cell Exhaustion Due to a Stage-Speci c Nonredundant Functional Relay Bowen Hou; ... et. al J Immunol (2024) 212 (3): 397–409. https://doi.org/10.4049/jimmunol.2300462 Related Content Smooth Muscle Cells Relay Acute Pulmonary In ammation via Distinct ADAM17/ErbB Axes J Immunol (January,2014) Crlz-1 Controls Germinal Center Reaction by Relaying a Wnt Signal to the Bcl-6 Expression in Centroblasts during Humoral Immune Responses J Immunol (November,2019) DNA Methylation Is a Nonredundant Repressor of the Th2 Effector Program J Immunol (October,2004) D ow nloaded from http://journals.aai.org/jim m unol/article-pdf/212/3/397/1651451/ji2300462.pdf by guest on 24 June 2025

Blocking Assay:

Image Search Results

SHP2 is identified as a PLCγ-dependent VEGFR2 pY1175 interaction partner in endothelial cells (A) Representative western blots showing VEGFR2 immunoprecipitated with antibodies against RASA1, CSK, VAV2, or SHP2 in HUVECs unstimulated (−) or stimulated (+) with VEGFA (100 ng/mL, 5 min). Corresponding whole-cell lysates analyzed using antibodies against phosphorylated VEGFR2 (pY1175), total VEGFR2, and GAPDH as loading control. (B) Quantification of VEGFR2 interaction with RASA1, CSK, VAV2, and SHP2 from (A); n = 3 independent experiments. (C) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCtr or siPTPN11. Scale bars: 30 μm. (D and E) Quantifications of MFI from (C), displayed as fold change relative to unstimulated control. (D) MFI of the VEC area. (E) MFI of pVEC Y685; n = 3 independent experiments, ≥3 fields of view per experiment. (F) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCSK or siVAV2. Scale bars: 30 μm. (G and H) Quantifications of MFI from (F), shown as fold change over unstimulated control; n = 3 independent experiments, ≥3 fields of view per experiment. (I) Representative western blot showing VEGFR2 immunoprecipitated with antibodies against SHP2 or IgG control from unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pre-treated with siCtr or siPLCG1 . Corresponding whole-cell lysates analyzed by blotting with antibodies against pVEGFR2 Y1175, VEGFR2, PLCγ, and GAPDH as loading control. (J) Quantification of VEGFR2-SHP2 binding from (I); n = 3 independent experiments. (K) Representative western blot showing downstream VEGFA-induced signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs, for 2, 5, 10, and 20 min pretreated with siCtr or siPTPN11. (L–N) Quantification of western blots from (K), shown as fold change relative to unstimulated control. (L) Quantification of western blots for peNOS S1177. (M) Quantification of western blots for pSFK Y418. (N) Quantification of western blots for pVEGFR2 Y1175; n = 4–5 independent experiments. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin; MFI, mean fluorescence intensity. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: SHP2 is identified as a PLCγ-dependent VEGFR2 pY1175 interaction partner in endothelial cells (A) Representative western blots showing VEGFR2 immunoprecipitated with antibodies against RASA1, CSK, VAV2, or SHP2 in HUVECs unstimulated (−) or stimulated (+) with VEGFA (100 ng/mL, 5 min). Corresponding whole-cell lysates analyzed using antibodies against phosphorylated VEGFR2 (pY1175), total VEGFR2, and GAPDH as loading control. (B) Quantification of VEGFR2 interaction with RASA1, CSK, VAV2, and SHP2 from (A); n = 3 independent experiments. (C) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCtr or siPTPN11. Scale bars: 30 μm. (D and E) Quantifications of MFI from (C), displayed as fold change relative to unstimulated control. (D) MFI of the VEC area. (E) MFI of pVEC Y685; n = 3 independent experiments, ≥3 fields of view per experiment. (F) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCSK or siVAV2. Scale bars: 30 μm. (G and H) Quantifications of MFI from (F), shown as fold change over unstimulated control; n = 3 independent experiments, ≥3 fields of view per experiment. (I) Representative western blot showing VEGFR2 immunoprecipitated with antibodies against SHP2 or IgG control from unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pre-treated with siCtr or siPLCG1 . Corresponding whole-cell lysates analyzed by blotting with antibodies against pVEGFR2 Y1175, VEGFR2, PLCγ, and GAPDH as loading control. (J) Quantification of VEGFR2-SHP2 binding from (I); n = 3 independent experiments. (K) Representative western blot showing downstream VEGFA-induced signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs, for 2, 5, 10, and 20 min pretreated with siCtr or siPTPN11. (L–N) Quantification of western blots from (K), shown as fold change relative to unstimulated control. (L) Quantification of western blots for peNOS S1177. (M) Quantification of western blots for pSFK Y418. (N) Quantification of western blots for pVEGFR2 Y1175; n = 4–5 independent experiments. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin; MFI, mean fluorescence intensity. See also .

Article Snippet: SHP2 chemical inhibition was performed by treating cells with 30 μM SHP099 (MedChemExpress, HY-100388) in Dimethyl sulfoxide (DMSO) diluted in starvation medium for 3 h. Starvation media with DMSO was used as the unstimulated control.

Techniques: Western Blot, Immunoprecipitation, Control, Binding Assay, Fluorescence

Endothelial PLCγ/SHP2 signaling mediates activation of Src by regulation of both the activating and inhibitory tyrosine phosphorylation sites (A) PLA using antibodies against Src and pSFK Y418 to detect phosphorylation of Src on Y418 in unstimulated or VEGFA-stimulated HUVECs (100 ng/mL, 5min), pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (B and C) MFI quantifications from (A), displayed as fold change relative to unstimulated control. (B) MFI of the total PLA signal. (C) MFI of the junctional PLA signals representing Y418 phosphorylation of Src; n = 7 independent experiments, ≥3 fields of view per experiment. (D) Representative western blot showing pSFK Y529 signaling in unstimulated (−) or VEGFA-stimulated HUVECs (100 ng/mL) for 2, 5, 10 and 20 min, pretreated with siCtr or si PTPN11 . (E) Quantification of western blots from (D); n = 5 independent experiments. (F and G) PLA using antibodies against Src and pSFK 529, visualizing phosphorylation of Src at the inhibitory phosphosite in HUVECs stimulated for 2, 5, and 10 min or left unstimulated. (F) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siCtr . (G) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at a higher magnification in panels below. Scale bars: 10 μm. (H) Quantification of PLA experiments from (F) and (G) and F; n = 6 independent experiments, ≥3 fields of view per experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. PLA, proximity ligation assay; HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: Endothelial PLCγ/SHP2 signaling mediates activation of Src by regulation of both the activating and inhibitory tyrosine phosphorylation sites (A) PLA using antibodies against Src and pSFK Y418 to detect phosphorylation of Src on Y418 in unstimulated or VEGFA-stimulated HUVECs (100 ng/mL, 5min), pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (B and C) MFI quantifications from (A), displayed as fold change relative to unstimulated control. (B) MFI of the total PLA signal. (C) MFI of the junctional PLA signals representing Y418 phosphorylation of Src; n = 7 independent experiments, ≥3 fields of view per experiment. (D) Representative western blot showing pSFK Y529 signaling in unstimulated (−) or VEGFA-stimulated HUVECs (100 ng/mL) for 2, 5, 10 and 20 min, pretreated with siCtr or si PTPN11 . (E) Quantification of western blots from (D); n = 5 independent experiments. (F and G) PLA using antibodies against Src and pSFK 529, visualizing phosphorylation of Src at the inhibitory phosphosite in HUVECs stimulated for 2, 5, and 10 min or left unstimulated. (F) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siCtr . (G) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at a higher magnification in panels below. Scale bars: 10 μm. (H) Quantification of PLA experiments from (F) and (G) and F; n = 6 independent experiments, ≥3 fields of view per experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. PLA, proximity ligation assay; HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin. See also .

Techniques: Activation Assay, Phospho-proteomics, Staining, Control, Western Blot, Proximity Ligation Assay

PLCγ/SHP2 interplay leads to eNOS activation followed by Src nitration (A) Representative western blot showing eNOS T495 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10 and 20 min, pre-treated with siCtr or siPTPN11 . (B) Quantification of western blots from (A); n = 4 independent experiments. (C) PLA for NitroTyr and pSFK Y418 to detect full activation of Src in HUVECs stimulated with VEGFA (100 ng/mL, 5 min) or left unstimulated, and pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (D) Quantification of junctional MFI PLA signals representing Y418 phosphorylation and 3-nitration of Src from (C), displayed as fold change to unstimulated control; n = 5 independent experiments, ≥3 fields of view per experiment. (E) Western blot showing eNOS S1177 and SHP2 Y542 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10, and 20 min, pre-treated with siCtr or siPLCG1 . (F and G) Quantifications of western blots from (E). (F) Quantifications of western blots for peNOS S1177. (G) Quantifications of western blots for pSHP2 Y542; n = 4 independent experiments. (H) Representative immunostaining images with antibodies against VE-cadherin (VEC; magenta) and pSHP2 Y542 (green), in HUVECs unstimulated or stimulated with VEGFA (100 ng/mL, 5min) after downregulation with siCtr or siPLCG1 . Scale bars: 30 μm. (I) Quantification of MFI from (H), shown as fold change relative to unstimulated control; n = 4 independent experiments, ≥3 fields of view/experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. VEC, VE-cadherin. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: PLCγ/SHP2 interplay leads to eNOS activation followed by Src nitration (A) Representative western blot showing eNOS T495 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10 and 20 min, pre-treated with siCtr or siPTPN11 . (B) Quantification of western blots from (A); n = 4 independent experiments. (C) PLA for NitroTyr and pSFK Y418 to detect full activation of Src in HUVECs stimulated with VEGFA (100 ng/mL, 5 min) or left unstimulated, and pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (D) Quantification of junctional MFI PLA signals representing Y418 phosphorylation and 3-nitration of Src from (C), displayed as fold change to unstimulated control; n = 5 independent experiments, ≥3 fields of view per experiment. (E) Western blot showing eNOS S1177 and SHP2 Y542 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10, and 20 min, pre-treated with siCtr or siPLCG1 . (F and G) Quantifications of western blots from (E). (F) Quantifications of western blots for peNOS S1177. (G) Quantifications of western blots for pSHP2 Y542; n = 4 independent experiments. (H) Representative immunostaining images with antibodies against VE-cadherin (VEC; magenta) and pSHP2 Y542 (green), in HUVECs unstimulated or stimulated with VEGFA (100 ng/mL, 5min) after downregulation with siCtr or siPLCG1 . Scale bars: 30 μm. (I) Quantification of MFI from (H), shown as fold change relative to unstimulated control; n = 4 independent experiments, ≥3 fields of view/experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. VEC, VE-cadherin. See also .

Techniques: Activation Assay, Nitration, Western Blot, Staining, Phospho-proteomics, Control, Immunostaining

VEGFR2 pY1173/PLCγ-induced vascular permeability in vivo requires SHP2 (A) Representative immunostaining images with antibodies against VEC (magenta) and pSHP2 Y542 (green) in the back skin of WT and Plcg1 iECKO mice after intradermal injection of PBS or VEGFA. Scale bars: 50 μm. (B) Quantification of MFI values from (A), for vascular pSHP2 Y542, displayed as fold change relative to PBS control; n = 5 mice/genotype, ≥3 fields of view/mouse. (C) Representative images of immunostaining for VEC (magenta) and pVEC Y685 (green) in the back skin of WT mice, intradermally injected with DMSO or SHP099 and subsequently PBS or VEGFA at the same site. Scale bars: 50 μm. (D) Quantification of MFI values from (C), for vascular pVEC Y685, shown as fold change relative to PBS control; n = 5 (DMSO) and 4 (SHP099) WT mice, ≥3 fields of view/mouse. (E) The Miles assay showing Evans blue leakage in the back skin of DMSO (control) or SHP099 treated WT mice, intradermally injected with PBS or VEGFA. (F) Quantification of extravasated Evans blue from (E), shown as fold change of DMSO-PBS-treated mice; n ≥ 8 mice/condition. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. VEC, VE-cadherin. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: VEGFR2 pY1173/PLCγ-induced vascular permeability in vivo requires SHP2 (A) Representative immunostaining images with antibodies against VEC (magenta) and pSHP2 Y542 (green) in the back skin of WT and Plcg1 iECKO mice after intradermal injection of PBS or VEGFA. Scale bars: 50 μm. (B) Quantification of MFI values from (A), for vascular pSHP2 Y542, displayed as fold change relative to PBS control; n = 5 mice/genotype, ≥3 fields of view/mouse. (C) Representative images of immunostaining for VEC (magenta) and pVEC Y685 (green) in the back skin of WT mice, intradermally injected with DMSO or SHP099 and subsequently PBS or VEGFA at the same site. Scale bars: 50 μm. (D) Quantification of MFI values from (C), for vascular pVEC Y685, shown as fold change relative to PBS control; n = 5 (DMSO) and 4 (SHP099) WT mice, ≥3 fields of view/mouse. (E) The Miles assay showing Evans blue leakage in the back skin of DMSO (control) or SHP099 treated WT mice, intradermally injected with PBS or VEGFA. (F) Quantification of extravasated Evans blue from (E), shown as fold change of DMSO-PBS-treated mice; n ≥ 8 mice/condition. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. VEC, VE-cadherin. See also .

Techniques: Permeability, In Vivo, Immunostaining, Injection, Control

VEGFR2 Y1173 heterozygosity is accompanied by decreased tumor endothelial PLCγ/SHP2 signaling and tumor vascular leakage (A) Representative immunostaining images of Vegfr2 +/+ (WT) and Vegfr2 Y1173F/+ (Y1173F/+) B16F10 melanoma tumors, showing vessels (IB4; red), pSHP2 Y542 (yellow), and fibrinogen (green). (B and C) Quantification of MFI from (A), shown as fold change over WT. (B) MFI for pSHP2 Y542. (C) MFI for fibrinogen; n = 5 (WT) and 3 (Y1173F/+) mice, ≥3 fields of view/mouse. (D) Representative immunostaining images of Vegfr2 +/+ (WT) and Vegfr2 Y1173F/+ (Y1173F/+) B16F10 melanoma tumors, showing vessels (CD31; red), pVEC Y731 (green), and CD45 (cyan). (E and F) Quantification of MFI from (D), displayed as fold change of WT. (E) MFI for pVEC Y731. (F) MFI for CD45 + cells (F); n = 5 (WT) and 3 (Y1173F/+), ≥3 fields of view/mouse. Scale bars: 100 μm. (G) Representative immunostaining images of WT and Plcg1 iECKO B16F10 melanoma tumors, showing vessels (IB4; red), pSHP2 Y542 (yellow), and fibrinogen (green). (H and I) MFI quantifications from (G), shown as fold change of WT. (H) MFI for pSHP2 Y542. (I) MFI for fibrinogen; n = 7 (WT) and 7 ( Plcg1 iECKO ), ≥3 fields of view/mouse. Unpaired 2-tailed Student’s t test. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. MFI, mean fluorescence intensity. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: VEGFR2 Y1173 heterozygosity is accompanied by decreased tumor endothelial PLCγ/SHP2 signaling and tumor vascular leakage (A) Representative immunostaining images of Vegfr2 +/+ (WT) and Vegfr2 Y1173F/+ (Y1173F/+) B16F10 melanoma tumors, showing vessels (IB4; red), pSHP2 Y542 (yellow), and fibrinogen (green). (B and C) Quantification of MFI from (A), shown as fold change over WT. (B) MFI for pSHP2 Y542. (C) MFI for fibrinogen; n = 5 (WT) and 3 (Y1173F/+) mice, ≥3 fields of view/mouse. (D) Representative immunostaining images of Vegfr2 +/+ (WT) and Vegfr2 Y1173F/+ (Y1173F/+) B16F10 melanoma tumors, showing vessels (CD31; red), pVEC Y731 (green), and CD45 (cyan). (E and F) Quantification of MFI from (D), displayed as fold change of WT. (E) MFI for pVEC Y731. (F) MFI for CD45 + cells (F); n = 5 (WT) and 3 (Y1173F/+), ≥3 fields of view/mouse. Scale bars: 100 μm. (G) Representative immunostaining images of WT and Plcg1 iECKO B16F10 melanoma tumors, showing vessels (IB4; red), pSHP2 Y542 (yellow), and fibrinogen (green). (H and I) MFI quantifications from (G), shown as fold change of WT. (H) MFI for pSHP2 Y542. (I) MFI for fibrinogen; n = 7 (WT) and 7 ( Plcg1 iECKO ), ≥3 fields of view/mouse. Unpaired 2-tailed Student’s t test. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. MFI, mean fluorescence intensity. See also .

Techniques: Immunostaining, Fluorescence

SHP2 and PLCγ are clinically relevant biomarkers for tumor vascular leakage (A and B) Representative multiplex images of RCC patient biopsies. (A) Multiplex images for RCC patients with low vascular PLCγ and SHP2 expression. (B) Multiplex images for RCC patients with high vascular PLCγ and SHP2 expression. Tissue sections were stained for PLCγ (red), SHP2 (yellow), CD34 (green), and FpA (white) to visualize vascular leakage, and counterstained with Hoesht. Scale bars: 100 μm. Boxed regions are shown at higher magnification to the right. Scale bars: 30 μm. (C) Heatmap comparing endothelial SHP2 expression, PLCγ expression, and vascular leakage scores across individual RCC patients ( n = 16). Patients are arranged by their ID number, with color coding indicating high (green), medium (yellow), or low (red) levels. (D) Schematic for the mechanism of SHP2/PLCγ interaction upon VEGFR2 activation. In summary, VEGFA stimuli leads to the phosphorylation of VEGFR2 Y1175, which recruits both PLCγ and SHP2. SHP2 is needed for the activation of PLCγ at the plasma membrane and hydrolysis of PIP2 to IP3 and DAG. The former triggers the release of intracellular calcium (Ca 2+ ), and together with DAG, activates PKC, causing phosphorylation of the eNOS activating site S1177 and dephosphorylation of the inactivating site T495. Production of NO mediates nitration and full activation of Src, which phosphorylates Y685 of VEC, disrupting the adherens junctions and resulting in vascular leakage. Created in BioRender. Kremmyda, P. (2025). RCC, renal cell carcinoma; FpA, fibrinopeptide A; PIP2, phosphatidylinositol-4,5-bisphosphate; IP3, inositol-1,4,5-trisphosphate; DAG, diacylglycerol; PKC, protein kinase C; eNOS, endothelial nitric oxide synthase; NO, nitric oxide; VEC, VE-cadherin. See also .

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: SHP2 and PLCγ are clinically relevant biomarkers for tumor vascular leakage (A and B) Representative multiplex images of RCC patient biopsies. (A) Multiplex images for RCC patients with low vascular PLCγ and SHP2 expression. (B) Multiplex images for RCC patients with high vascular PLCγ and SHP2 expression. Tissue sections were stained for PLCγ (red), SHP2 (yellow), CD34 (green), and FpA (white) to visualize vascular leakage, and counterstained with Hoesht. Scale bars: 100 μm. Boxed regions are shown at higher magnification to the right. Scale bars: 30 μm. (C) Heatmap comparing endothelial SHP2 expression, PLCγ expression, and vascular leakage scores across individual RCC patients ( n = 16). Patients are arranged by their ID number, with color coding indicating high (green), medium (yellow), or low (red) levels. (D) Schematic for the mechanism of SHP2/PLCγ interaction upon VEGFR2 activation. In summary, VEGFA stimuli leads to the phosphorylation of VEGFR2 Y1175, which recruits both PLCγ and SHP2. SHP2 is needed for the activation of PLCγ at the plasma membrane and hydrolysis of PIP2 to IP3 and DAG. The former triggers the release of intracellular calcium (Ca 2+ ), and together with DAG, activates PKC, causing phosphorylation of the eNOS activating site S1177 and dephosphorylation of the inactivating site T495. Production of NO mediates nitration and full activation of Src, which phosphorylates Y685 of VEC, disrupting the adherens junctions and resulting in vascular leakage. Created in BioRender. Kremmyda, P. (2025). RCC, renal cell carcinoma; FpA, fibrinopeptide A; PIP2, phosphatidylinositol-4,5-bisphosphate; IP3, inositol-1,4,5-trisphosphate; DAG, diacylglycerol; PKC, protein kinase C; eNOS, endothelial nitric oxide synthase; NO, nitric oxide; VEC, VE-cadherin. See also .

Techniques: Multiplex Assay, Expressing, Staining, Activation Assay, Phospho-proteomics, Clinical Proteomics, Membrane, De-Phosphorylation Assay, Nitration

Journal: Redox Biology

Article Title: Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis

doi: 10.1016/j.redox.2026.104096

Figure Lengend Snippet: HDCA modulates Treg migration and atherosclerotic plaque composition via FXR signaling. ApoE−/− mice (C57BL/6J background, male, 8 weeks old) were fed a high-fat diet for 28 days to induce AS and subsequently received adoptive transfer of control or FXR-knockout (FXR KO) Treg cells generated by CRISPR/Cas9-mediated lentiviral transduction, with HDCA (30 μM) or vehicle treatment as indicated. (A) Representative Western blot analysis of FXR, PD-1, SHP-2, p-Raptor, RAC and IL-10R expression in isolated Treg cells from each group. (B) Oil Red O staining was performed to assess lipid accumulation in the aorta. (C) Masson's trichrome staining of aortic sections from FXR KO mice reveals comparable plaque area and collagen deposition in both HDCA-treated and untreated groups (magnification, 5 × ; scale bar, 1 mm). (D) Representative H&E images of aortic sections show the difference between untreated and HDCA-treated mice in the FXR KO groups (magnification, 5 × ; scale bar, 500 μm). Relative bar graphs show quantification of lesion area and lesion/media area ratio. (E) Confocal immunofluorescence was used to evaluate Foxp3+ Treg infiltration within atherosclerotic plaques (scale bar, 25 μm). (F) Immunohistochemistry images show Treg accumulation in the plaque area across all groups (magnification, 40 × ; scale bar, 100 μm). (G) Flow cytometry analysis of Treg proportions in aortic plaques and spleen from control and FXR KO mice, with or without HDCA treatment. (H) Western blot analysis of matrix remodeling-related proteins, including calpain 1 and matrix metalloproteinase 2, and the anti-inflammatory factor IL-10. Data are presented as mean ± SD (n = 5 biological replicates). Data with four groups were analyzed by one-way ANOVA with Tukey's post hoc test. Comparisons between two groups were performed using the non-parametric Mann-Whitney U test. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Proteins were detected using the following antibodies: anti-CPT1a antibody (ab234111, abcam), anti-beta actin antibody (ab8226, abcam), anti-PERK antibody (ab229912, abcam), anti-ERK1+ERK2 antibody (ab184699, abcam), anti-S6K1 antibody (ab14708, abcam), anti-S6K1 (phospho T229) antibody (ab5231, abcam), Rac1/2/3 antibody (G-2) (sc-514583, Santa Cruz), anti-Calpain 1 antibody (ab108400, abcam), anti-MMP2 antibody (ab92536, abcam), anti-IL-10 antibody (ab310329, abcam), anti-ZNF671 antibody (HPA046099, Sigma-Aldrich), anti-MAPK6/ERK3 antibody (ab53277, abcam), SIAH1 recombinant rabbit monoclonal antibody (PSH01-80) (MA5-51926, Thermo Fisher), p-Stat1 antibody (pY701.4A) (sc-136229, Santa Cruz), Stat1 antibody (C-136) (sc-464, Santa Cruz), anti-FXR1 antibody (ab155124, abcam), phospho-Raptor (Ser792) polyclonal antibody (PA5-118730, Thermo Fisher), anti-PD1 antibody (ab214421, abcam), SHP-2 antibody (3752S, Cell Signaling Technology), IL-10R antibody (3F9) (sc-53654, Santa Cruz), GAPDH antibody (6C5) (sc-32233, Santa Cruz), rabbit anti-mouse IgG H&L (HRP) (ab6728, abcam), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody, Alexa FluorTM Plus 488 (A32731, Thermo Fisher).

Techniques: Migration, Adoptive Transfer Assay, Control, Knock-Out, Generated, CRISPR, Transduction, Western Blot, Expressing, Isolation, Staining, Immunofluorescence, Immunohistochemistry, Flow Cytometry, MANN-WHITNEY

NFIC promotes OGN and PTEN expression while inhibiting NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot analysis of NFIC, OGN, p-NF-κB, SHP2, and p-SHP2 protein bands in six groups (NFIC-NC, NFIC-OE, NF-κB NC, NF-κB NC, PHPS1 NC, PHPS1 OE), with statistical analysis of relative protein expression levels. B: Western blot analysis of PTEN and HEY1 protein bands in six groups, along with statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while inhibiting NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot analysis of NFIC, OGN, p-NF-κB, SHP2, and p-SHP2 protein bands in six groups (NFIC-NC, NFIC-OE, NF-κB NC, NF-κB NC, PHPS1 NC, PHPS1 OE), with statistical analysis of relative protein expression levels. B: Western blot analysis of PTEN and HEY1 protein bands in six groups, along with statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Expressing, Western Blot, Standard Deviation

NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot detection of p-PIK, p-AKT, p-STAT3, GAPDH protein bands, and statistical analysis of relative protein expression levels; B: Western blot detection of Cyclin A1, Cyclin D1, MMP-3, and MMP-9 protein bands in the six groups, and statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot detection of p-PIK, p-AKT, p-STAT3, GAPDH protein bands, and statistical analysis of relative protein expression levels; B: Western blot detection of Cyclin A1, Cyclin D1, MMP-3, and MMP-9 protein bands in the six groups, and statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Techniques: Expressing, Western Blot, Standard Deviation

NFIC inhibits glioblastoma cell proliferation and invasion, while NF-κB promotes these processes. A: Co-immunoprecipitation (CO-IP) analysis of NFIC and PTEN protein bands in six sample groups; B: CO-IP analysis of OGN and NF-κB protein bands in six sample groups; C: CO-IP analysis of NF-κB and SHP2 protein bands in six sample groups; D: CO-IP analysis of NFIC and OGN protein bands in six sample groups; Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference; * P < 0.05; ** P < 0.01; nsP > 0.05.

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC inhibits glioblastoma cell proliferation and invasion, while NF-κB promotes these processes. A: Co-immunoprecipitation (CO-IP) analysis of NFIC and PTEN protein bands in six sample groups; B: CO-IP analysis of OGN and NF-κB protein bands in six sample groups; C: CO-IP analysis of NF-κB and SHP2 protein bands in six sample groups; D: CO-IP analysis of NFIC and OGN protein bands in six sample groups; Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference; * P < 0.05; ** P < 0.01; nsP > 0.05.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Standard Deviation

NFIC binds to the promoter regions of OGN and PTEN and regulates their transcription, leading to increased expression of these two genes. Immunohistochemical staining results for NFIC, PTEN, OGN, NF-κB, and p-SHP2, along with statistical analysis of staining. Data are presented as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, *P < 0.05, **P < 0.01.

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC binds to the promoter regions of OGN and PTEN and regulates their transcription, leading to increased expression of these two genes. Immunohistochemical staining results for NFIC, PTEN, OGN, NF-κB, and p-SHP2, along with statistical analysis of staining. Data are presented as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, *P < 0.05, **P < 0.01.

Techniques: Expressing, Immunohistochemical staining, Staining, Standard Deviation

NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, p-SHP2, PI3K, AKT, Cyclin A1, Cyclin D1, MMP-3, and MMP-9 expression. NF-κB promotes SHP2 expression, while OGN and PTEN inhibit p-SHP2 expression. NFIC suppresses glioma cell proliferation and invasion, whereas NF-κB promotes these processes.

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, p-SHP2, PI3K, AKT, Cyclin A1, Cyclin D1, MMP-3, and MMP-9 expression. NF-κB promotes SHP2 expression, while OGN and PTEN inhibit p-SHP2 expression. NFIC suppresses glioma cell proliferation and invasion, whereas NF-κB promotes these processes.

Techniques: Expressing

Review

shp 2 antibody (Cell Signaling Technology Inc)

Structured Review

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