shp 2 antibody  (Cell Signaling Technology Inc)

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

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

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.

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

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.

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: Immunoprecipitation, Co-Immunoprecipitation Assay, Standard Deviation

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.

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, Immunohistochemical staining, Staining, Standard Deviation

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.

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

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 .

Article Snippet: Primary antibodies diluted 1:100 in antibody diluent and added for overnight incubation at 4°C include rabbit anti-pSHP2 (Y542) (Cell Signaling, 3751S, 1:250) and mouse anti-PLCγ (E−12) (SantaCruz, sc-7290), rabbit anti–phospho-SFK (Tyr416) (Cell Signaling Technology, 2101s), rabbit anti-phospho-Src (Tyr527) (Cell Signaling Technology, 2105s), mouse anti-Src (Abcam, ab231081), mouse anti-Yes (BD Biosciences, 610376), and mouse anti-3-Nitrotyrosine (Abcam, ab61392).

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

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 .

Article Snippet: Primary antibodies diluted 1:100 in antibody diluent and added for overnight incubation at 4°C include rabbit anti-pSHP2 (Y542) (Cell Signaling, 3751S, 1:250) and mouse anti-PLCγ (E−12) (SantaCruz, sc-7290), rabbit anti–phospho-SFK (Tyr416) (Cell Signaling Technology, 2101s), rabbit anti-phospho-Src (Tyr527) (Cell Signaling Technology, 2105s), mouse anti-Src (Abcam, ab231081), mouse anti-Yes (BD Biosciences, 610376), and mouse anti-3-Nitrotyrosine (Abcam, ab61392).

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

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 .

Article Snippet: Primary antibodies diluted 1:100 in antibody diluent and added for overnight incubation at 4°C include rabbit anti-pSHP2 (Y542) (Cell Signaling, 3751S, 1:250) and mouse anti-PLCγ (E−12) (SantaCruz, sc-7290), rabbit anti–phospho-SFK (Tyr416) (Cell Signaling Technology, 2101s), rabbit anti-phospho-Src (Tyr527) (Cell Signaling Technology, 2105s), mouse anti-Src (Abcam, ab231081), mouse anti-Yes (BD Biosciences, 610376), and mouse anti-3-Nitrotyrosine (Abcam, ab61392).

Techniques: Immunostaining, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Antitumor effects of LPM5140276 and its potential combination with SHP2 inhibition in KRAS G12D -mutant cancer

doi: 10.3389/fphar.2025.1554356

Figure Lengend Snippet: Effects of LPM5140276 and/or RMC4550 on the signaling pathways related to cell proliferation, cell-cycle arrest, and apoptosis. (A) In the AsPC-1 cells, p-ERK and p-AKT levels were reduced in a dose-dependent manner 3 h after LPM5140276 treatment. (B) LPM5140276-treated AsPC-1 cells showed downregulated CDK4 and cyclin D1 expression at 3 h, 12 h, 24 h, and 48 h. (C) p-ERK levels decreased from 3 h to 48 h but rebounded at 72 h, while p-AKT levels decreased from 3 h to 24 h and rebounded at 48 h. LPM5140276 treatment upregulated cleaved caspase-3 and cleaved caspase-7 levels from 3 h to 72 h in the AsPC-1 cells. (D) LPM5140276 monotherapy reduced p-ERK level in a concentration-dependent manner but increased p-SHP2 level in the AsPC-1 cells at 3 h. Compared to single-agent treatments, combined administration of RMC4550 and LPM5140276 at increasing concentrations resulted in greater suppression of both p-ERK and p-SHP2 at 3 h.

Article Snippet: Antibodies against Ras (G12D-mutant specific; #14429), SHP2 (#3752), p-SHP2 (#3751), ERK1/2 (#4695), p-ERK1/2 (#4370), AKT (#4691), p-AKT (#4058), CDK4 (#12790), cyclin D1 (#2922), cleaved caspase-3 (#9661), and cleaved caspase-7 (#9491) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Expressing, Concentration Assay