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

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 .

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

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: 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: 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: 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: 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: 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: Immunostaining, Fluorescence

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 .

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

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 1. Decreased ethanol-induced hepatic injury and inflammation in mice with Shp2 pharmacological inhibition. Wild-type female mice were subjected to the chronic plus binge model without (DMSO) and with Shp2 pharmacological inhibitor (Shp2i). (A) Body weight (%), (B) plasma ALT level, (C) hepatic mRNA expression of Ccl2, Tnfa, Il1b, Adgre1, Ly6g, and Cd68, and (D) immunoblots of hepatic pNF-κB p65 S536 and NF-κB p65 from mice fed without (Pair) and with ethanol (EtOH) diet. Expression fold change of target mRNAs was normalized to Tbp. The phosphorylation level of NF-κB p65 was normalized to NF-κB p65 and summarized in a dot plot. Each lane in the immunoblot represents a sample from an individual mouse. Sample numbers for (B), (C), and (D) are indicated in the panels and n=6–10 in (A). A.U.: arbitrary unit. *p < 0.05, **p < 0.01 Pair versus EtOH of mice with the same intervention. †p < 0.05, ††p < 0.01 DMSO versus Shp2i of mice with the same diet.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Inhibition, Clinical Proteomics, Expressing, Western Blot, Phospho-proteomics

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 2. Shp2 pharmacological inhibition mitigated ethanol-induced hepatic steatosis. Hepatic (A) triglycerides, (B) mRNA of Pparg, Cd36, Srebf1, Ppara, and Scd1, and (C) H&E staining from mice fed without (Pair) and with ethanol (EtOH) diet and co-treated without (DMSO) and with Shp2 inhibitor (Shp2i). Sample numbers for (A) and (B) are indicated in the panels. Expression fold change of target mRNAs was normalized to Tbp. A.U.: arbitrary unit. Scale bar: 100 µm. Representative H&E images from three individual mice per group are presented. Statistics for (A), *p < 0.05 DMSO versus Shp2i of mice with the EtOH diet in the triglycerides assay; and for (B), **p < 0.01 Pair versus EtOH of mice with the same intervention, and ††p < 0.01 DMSO versus Shp2i of mice with the same diet.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Inhibition, Staining, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 3. Decreased ethanol-induced ERK activation, oxidative, and ER stress in the livers of mice with Shp2 inactivation. Immunoblots of hepatic lysates for (A) CYP2E1, ADH1, ALDH1/2, GAPDH, (B) pERK T202/Y204, ERK, (C) iNOS, 4HNE, Actin, and (D) pPERK T981, PERK, peIF2α S51, eIF2α, spliced XBP1 (sXBP1), CHOP, and GAPDH in mice fed without (Pair) and with ethanol (EtOH) diet and co-treated without (DMSO) and with Shp2 inhibitor (Shp2i). Each lane in the immunoblot represents a sample from an individual mouse. Quantification results were summarized in dot plots with relevant normalization as indicated. Sample numbers for (B), (C), and (D) are indicated in the panels. A.U.: arbitrary unit. **p < 0.01 Pair versus EtOH of mice with the same intervention. †p < 0.05, ††p < 0.01 DMSO versus Shp2i of mice with the same diet.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Activation Assay, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 4. Diminished ethanol-evoked inflammation in the ileum of mice with Shp2 pharmacological inhibition. Wild-type female mice were subjected to the chronic plus binge model without (DMSO) and with Shp2 inhibition (Shp2i). (A) mRNA expressions of Il1b, Ccl2, Tnfa, Adgre1, Ly6g, and (B) immunoblots of pNF-κB p65 S536 and NF-κB p65 in ileum samples from mice fed without (Pair) and with ethanol (EtOH) diet. Expression fold change of target mRNAs was normalized to Tbp. The phosphorylation level of NF-κB p65 was normalized to NF-κB p65 and summarized in a dot plot. Each lane in the immunoblot represents a sample from an individual mouse. Sample numbers for (A) and (B) are indicated in the panels. A.U.: arbitrary unit. *p < 0.05, **p < 0.01 Pair versus EtOH of mice with the same intervention. †p < 0.05, ††p < 0.01 DMSO versus Shp2i of mice with the same diet.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Inhibition, Western Blot, Expressing, Phospho-proteomics

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 5. Shp2 inhibition ameliorated the ethanol-induced intestinal permeability in vivo and the reduction in the expression of tight junction proteins. (A) Intestinal permeability in vivo from ethanol-fed mice treated without (DMSO, n=6) and with Shp2 inhibition (Shp2i, n=10). *p < 0.05 DMSO versus Shp2i. (B) Blood endotoxin level and ileum (C) mRNA expression, (D) immunoblots, and (E) immunostainings of tight junction proteins occludin and ZO-1 from mice fed without (Pair) and with ethanol (EtOH) diet and co-treated with DMSO and Shp2i. Expression fold change of target mRNAs was normalized to Tbp. Each lane in the immunoblot represents a sample from an individual mouse. Sample numbers for (B) and (C) are indicated in the panels. Representative images of immunofluo rescence staining from three individual mice per group are presented. **p < 0.01 Pair versus EtOH of mice with the same intervention. †p < 0.05, ††p < 0.01 DMSO versus Shp2i of mice with the same diet. A.U.: arbitrary unit. Scale bar: 50 µm.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Inhibition, Permeability, In Vivo, Expressing, Western Blot, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Pharmacological inhibition of the Src homology phosphatase 2 confers partial protection in a mouse model of alcohol-associated liver disease.

doi: 10.1016/j.biopha.2024.116590

Figure Lengend Snippet: Fig. 6. Shp2 inhibition attenuated ethanol-induced ERK and Src activation and oxidative and ER stress in the ileum of mice. Ileum lysates were immu noblotted for (A) pERK T202/Y204, ERK, pSrc Y416, Src, (B) iNOS, 4HNE, GAPDH, and (C) spliced XBP1 (sXBP1), cleaved Caspase3 (cCasp3), and GAPDH in mice fed without (Pair) and with ethanol (EtOH) diet and co-treated without (DMSO) and with Shp2 inhibitor (Shp2i). Each lane in the immunoblot represents a sample from an individual mouse. Quantification results were summarized in dot plots with relevant normalization as indicated. Sample numbers for (A), (B), and (C) are indicated in the panels. A.U.: arbitrary unit. *p < 0.05, **p < 0.01 Pair versus EtOH of mice with the same intervention. †p < 0.05, ††p < 0.01 DMSO versus Shp2i of mice with the same diet.

Article Snippet: On day eleven, mice obtained a single bolus of ethanol (5 g/kg body weight) or isocaloric maltose dextrin solution by oral gavage and sacrificed after 9 h. For Shp2 pharmacological inhibition, mice were intraperitoneally injected daily with 5 mg/kg of SHP099 (S8278, Selleck Chemicals; dissolved in DMSO) at the initiation of Pair- and EtOH-feeding.

Techniques: Inhibition, Activation Assay, Western Blot