Journal: Scientific Reports

Article Title: CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

doi: 10.1038/s41598-025-31475-3

Figure Lengend Snippet: Bioinformatics analysis reveals the crucial role of the CBX2-NF-κB-METTL3-SHP2 pathway in oral squamous cell carcinoma. ( A ) Boxplot of sample correction; ( B ) PCA plot of differential samples; ( C ) Volcano plot of differential genes; ( D ) Ordered plot of differential genes; ( E ) Bar chart of GO enrichment analysis; ( F ) Bubble plot of KEGG enrichment analysis; ( G ) Lollipop chart of KEGG enrichment analysis; ( H ) Scatter plot of CBX2 and CEP55 correlation; ( I ) Scatter plot of CEP55 and NF-κB correlation; ( J ) Scatter plot of NF-κB and METTL3 correlation; ( K ) Scatter plot of METTL3 and SHP2 (PTPN11) correlation; ( L ) CBX2 expression levels and patient survival analysis.

Article Snippet: NF-κB activator E1351 and SHP2 inhibitor PHPS1 were purchased from MedChemExpress, USA.

Techniques: Expressing

Journal: Scientific Reports

Article Title: CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

doi: 10.1038/s41598-025-31475-3

Figure Lengend Snippet: Western blot and Co-IP results indicate that knockdown of CBX2, activation of NF-κB, inhibition of METTL3, overexpression of METTL3, and Inhibition of SHP2 all upregulate or downregulate downstream signaling molecules. ( A ) Western Blot analysis of protein bands for CBX2, CEP55, p-NF-κB, METTL3, TGFβ1, and p-SHP2 in the NC group, CBX2-KD group, CBX2-KD-E1351 group, CBX2-KD-E1351-STM2457 group, CBX2-KD-E1351-STM2457-METTL3-OE group, and CBX2-KD-E1351-STM2457-METTL3-OE-PHPS1 group, as well as statistical analysis graphs of relative protein expression levels. GAPDH was as a control protein; ( B ) Western Blot analysis of protein bands for p-PI3K, Slug, and Snail, along with statistical analysis graphs of relative protein expression levels. GAPDH was as the control protein; ( C ) Statistical analysis chart of the Co-IP experiment analyzing the interaction between endogenous CBX2 and CEP55 in SCC-25 cells. N = 6; Data are expressed as mean ± standard deviation; * P < 0.05; ** P < 0.01; ns P > 0.05.

Article Snippet: NF-κB activator E1351 and SHP2 inhibitor PHPS1 were purchased from MedChemExpress, USA.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Knockdown, Activation Assay, Inhibition, Over Expression, Expressing, Control, Standard Deviation

Journal: Scientific Reports

Article Title: CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

doi: 10.1038/s41598-025-31475-3

Figure Lengend Snippet: Scratch, Transwell, and Colony Formation Experiments Indicate That Knockdown of CBX2, Activation of NF-κB, Inhibition of METTL3, Overexpression of METTL3, and Inhibition of SHP2 All Inhibit or Promote Tumor Cell Migration, Invasion, and Proliferation. ( A ) Analysis of scratch test results for NC group, CBX2-KD group, CBX2-KD-E1351 group, CBX2-KD-E1351-STM2457 group, CBX2-KD-E1351-STM2457-METTL3-OE group, and CBX2-KD-E1351-STM2457-METTL3-OE-PHPS1 group at 0 hours and 48 hours, as well as statistical analysis of scratch distance; ( B ) Analysis of Transwell experiment for SCC-25 cells, including the experimental results images for cell migration and invasion, as well as statistical analysis charts of the number of migrating and invading cells; ( C ) Colony formation analysis of the experimental results graph showing the proliferation of SCC-25 cells in experiments, as well as the statistical analysis graph of the number of clone cells. N=6; Data are expressed as mean ± standard deviation; * P < 0.05; ** P < 0.01; ns P > 0.05.

Article Snippet: NF-κB activator E1351 and SHP2 inhibitor PHPS1 were purchased from MedChemExpress, USA.

Techniques: Knockdown, Activation Assay, Inhibition, Over Expression, Migration, Standard Deviation

Journal: Scientific Reports

Article Title: CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

doi: 10.1038/s41598-025-31475-3

Figure Lengend Snippet: Western blot and tubulogenesis experiment results show that knockdown of CBX2, activation of NF-κB, inhibition of METTL3, overexpression of METTL3, and inhibition of SHP2 all inhibit or promote angiogenesis. ( A ) Western Blot analysis of VEGFA and HIF1α protein bands in the NC group, CBX2-KD group, CBX2-KD-E1351 group, CBX2-KD-E1351-STM2457 group, CBX2-KD-E1351-STM2457-METTL3-OE group, and CBX2-KD-E1351-STM2457-METTL3-OE-PHPS1 group, along with statistical analysis graphs of relative protein expression levels. GAPDH are as the control protein; ( B ) Tubulogenesis analysis of experimental results, including figures showing the formation of tube-like structures and statistical analysis of the number of neovessels per visual field. N=6; Data are expressed as mean ± standard deviation; * P < 0.05; ** P < 0.01; ns P > 0.05.

Article Snippet: NF-κB activator E1351 and SHP2 inhibitor PHPS1 were purchased from MedChemExpress, USA.

Techniques: Western Blot, Knockdown, Activation Assay, Inhibition, Over Expression, Expressing, Control, Standard Deviation

Journal: Scientific Reports

Article Title: CBX2 promoted oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling induced metastasis/proliferation and angiogenesis

doi: 10.1038/s41598-025-31475-3

Figure Lengend Snippet: Schematic illustration of CBX2 promoting oral squamous cell carcinoma via increasing CEP55/NF-κB/METTL3/SHP2 signaling, inducing metastasis/proliferation, and angiogenesis.

Article Snippet: NF-κB activator E1351 and SHP2 inhibitor PHPS1 were purchased from MedChemExpress, USA.

Techniques:

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: (i) Representative blots of Shp1 and Shp2 expression levels from CT, Shp1, Shp2 and Shp1/2 DKO mice by capillary immunoassays with the respective antibodies in (A) sorted-MK progenitors and (B) washed platelets. Percentage of residual level of (ii) Shp1 and (iii) Shp2 in Shp1 KO (blue), Shp2 KO (green) and Shp1/2 DKO (red) mice. n = 3-6 mice per genotype. Mean ± SEM; one way-ANOVA; ** P < 0.01, *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Expressing

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: (A) Platelet counts (i) and platelet volumes (ii) of control (CT) (n=75), Shp1 KO (n=21), Shp2 KO (n=29) and Shp1/2 DKO (n=46) mice. Mean ± SEM; one way-ANOVA; *** P < 0.001. (B) (i) Cumulative bleeding time was recorded (ii) and the volume of blood loss was measured over 30 minutes. Mean ± SEM; one way-ANOVA; * P < 0.05. (C) Mean fluorescence intensity (MFI) of platelet (i) GPVI and (ii) α2 integrin expression in CT (n=10), Shp1 KO (n=4), Shp2 KO (n=4) and Shp1/2 DKO (n=13) mice. Mean ± SEM; one way-ANOVA; *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Control, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: (A-C) Aggregation of washed platelets were measured by lumi-aggregometry in response to agonists indicated. Representative traces, n= 4-8 mice/genotype per condition, percentage of aggregation at 5 minutes and area under the curve (AUC) quantification. Mean ± SEM, n = 5-17 mice/genotype per condition, one way-ANOVA, * P < 0.05. (D) Mean fluorescence intensity (MFI) of P-selectin expression of control (CT), Shp1 KO, Shp2 KO and Shp1/2 DKO platelets in whole blood in response to (i) 1 and 3 μg/mL CRP and (ii) 100 and 500 μM PAR4 peptide (PAR4p). Mean ± SEM, n = 5-9 mice/genotype per condition, two way-ANOVA; * P < 0.05, *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Fluorescence, Expressing, Control

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: Whole cell lysates of resting and (A) 10 µg/ml CRP-stimulated and (B) 10 µg/ml activating CLEC-2 antibody platelets from Shp1 KO, Shp2 KO and Shp1/2 DKO mice and litter-matched CT mice were western blotted with anti-Src p-Ty418, Src total, -Syk p-Tyr519/520 and Syk antibodies. (i) Representative blots of capillary-based immunoassays and (ii) quantification of peak areas from three independent experiments, Mean ± SEM, n = 3-4 mice/genotype; one-way ANOVA, *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: (A) Mature BM-derived MKs from Shp1 KO, Shp2 KO, Shp1/2 DKO, and litter-matched CT mice were stained with propidium iodide and ploidy of cells was quantified by flow cytometry. (i-ii) The percentage of 2-4N and 8-128N ploidy cells was quantified (n = 4-6 mice/genotype; mean ± SEM; two-way ANOVA, * P < 0.05, ** P < 0.01, *** P < 0.001). (B) Ex vivo proplatelet formation. Percentage of MKs forming proplatelet was quantified in culture. Mean ± SEM, two-way ANOVA, *** P < 0.001. (C) Mature BM-derived MKs from Shp1 KO, Shp2 KO, Shp1/2 DKO and litter-matched CT mice were stimulated with 50 ng/mL thrombopoietin (Tpo) for 10 min at 37°C. Whole cell lysates were western blotted with indicated antibodies. Representative blots capillary-based immunoassays and quantification of peak areas from n = 3 independent experiments/genotype; two-way ANOVA, *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Derivative Assay, Staining, Flow Cytometry, Ex Vivo, Western Blot

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: Bone marrow explants. Proportion of MKs from Shp1 KO, Shp2 KO, Shp1/2 DKO and litter-matched CT mice extending proplatelets at 6h of observation were observed. Bars represent the mean ± SEM of six independent experiments. (A) Quantification and (B) representatives’ images of round MKs, MKs with large ends and proplatelet (PPT) forming MKs. Scale bar: 50 µm. Mean ± SEM, one-way ANOVA, * P < 0.1, *** P < 0.001. (C) Classification of the MK according to their maturation stage: stage I (absence of granules), stage II (granules and developing demarcation membrane system (DMS) not yet organized), stage III (DMS organized in cytoplasmic territories). Data are reported as the percentage of the total number of MK, (i) of all stage MK and (ii) only stage 3 MK. Bars represent the mean ± SEM in three BM samples, *** P < 0.001. (D) Ploidy of in situ BM MKs measured by flow cytometry. Mean ± SEM, two-way ANOVA, * P < 0.1, *** P < 0.001.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Membrane, In Situ, Flow Cytometry

Journal: bioRxiv

Article Title: Synergistic effects of deleting the tyrosine phosphatases Shp1 and Shp2 on megakaryopoiesis and thrombopoiesis in mice

doi: 10.1101/2025.10.24.684367

Figure Lengend Snippet: (A) Effects of two selective Shp1 allosteric inhibitors 10 µM M029 and 10 µM F2Ac were tested on megakaryopoeisis. (i) Viability, (ii) proliferation, and (iii) MK ploidy. Quantification from n = 3-4 independent experiments/condition. (B) Whole cell lysates of resting and 50 ng/mL Tpo-stimulated MKs were western blotted with the indicated antibodies. Representative blots capillary-based immunoassays and quantification of peak areas from n = 3 independent experiments/genotype. (C) Effects of two selective Shp2 allosteric inhibitors 10 µM SHP099 and 10 µM RMC-4550 were tested on megakaryopoeisis. (i) Viability, (ii) proliferation, (iii) MK ploidy and (iv) percentage of MKs forming proplatelets. Quantification from n = 3-6 independent experiments/condition. * P < 0.05; ** P < 0.01; *** P < 0.001, two-way ANOVA. (D) Whole cell lysates of resting and 50 ng/mL Tpo-stimulated MKs were western blotted with the indicated antibodies. Representative blots capillary-based immunoassays and quantification of peak areas from n = 3 independent experiments/genotype. * P < 0.05; ** P < 0.01; two-way ANOVA.

Article Snippet: SHP099 and RMC-4550 Shp2 allosteric inhibitors were purchased from MedChemExpress.

Techniques: Western Blot