Journal: JCI Insight

Article Title: COVID-19 and RA share an SPP1 myeloid pathway that drives PD-L1 + neutrophils and CD14 + monocytes

doi: 10.1172/jci.insight.147413

Figure Lengend Snippet: ( A ) Venn diagram illustrating numbers of unique and shared marker genes of ST TREM2 hi and BALF FABP4 + macrophage clusters as described in . Marker genes were identified prior to integration of data sets ( , ) and were calculated using MAST, setting a minimum percentage of cells in clusters expressing each marker to 40%. Genes considered differentially expressed at P < 0.05 after Bonferroni correction. ( B ) Heatmap illustrating scaled, pseudobulk expression of shared upregulated marker genes from ST and BALF clusters indicated in A . ( C ) Split UMAP plots comparing BALF macrophage clusters in health, and in mild and severe COVID-19, illustrating changes in expression of the TAM receptors AXL and MerTK , with their respective preferred ligands GAS6 and PROS1 . Intensity of purple indicates expression level. ( D ) Heatmap illustrating scaled, pseudobulk expression of TAM receptors and associated ligands by each BALF cluster, across patient groups. TAM receptors and their ligands were significantly differentially expressed in severe COVID-19 versus healthy tissues ( P ≤ 0.005), with Bonferroni correction for multiple comparison, as confirmed by MAST.

Article Snippet: Blood samples were centrifuged (600 g /15 minutes) and plasma aliquots were stored at –80°C until analysis by ELISA for SPP1 (BMS2066; Thermo Fisher Scientific), S100A12 (DY1052; R&D Systems), GAS6 (DY885B; R&D Systems), and PROS1 (NBP2-60585; NOVUS Biological).

Techniques: Marker, Expressing, Comparison

Journal: JCI Insight

Article Title: COVID-19 and RA share an SPP1 myeloid pathway that drives PD-L1 + neutrophils and CD14 + monocytes

doi: 10.1172/jci.insight.147413

Figure Lengend Snippet: ( A ) Patients and healthy donors, shown as the following: n = 121 patients with acute pneumonia ( n = 29 community acquired SARS-CoV-2 – pneumonia, n = 29 mild/moderate COVID-19, n = 63 severe COVID-19), convalescent COVID-19 ( n = 41), and healthy controls ( n = 10). Representative images of lung CT scans. ( B ) Plasma levels of SPP1, S100A12, GAS6, and PROS1 in groups as in A . ( C ) Spearman’s rank correlations between SPP1, S100A12, GAS6, and PROS1 plasma levels in patients with acute COVID-19 pneumonia ( n = 92) with demographic and clinical parameters. Each box displays the r value, and an asterisk indicates statistical significance of P < 0.05. ( D ) Plasma levels of SPP1, S100A12, GAS6, and PROS1 in patients with acute COVID-19 pneumonia ( n = 92) stratified based on lung functions measured by PaO 2 /FiO 2 at the time of hospital admission. Severe respiratory failure was defined by PaO 2 /FiO 2 ≤ 200. ( E ) Percentage of acute COVID-19 pneumonia patients ( n = 92) with PaO 2 /FiO 2 ≤ 200 based on high plasma levels of SPP1 (≥108 ng/mL), S100A12 (≥59 ng/mL), GAS6 (≥24 ng/mL), and PROS1 (≥15 μg/mL). ( F ) COVID-19 patient plasma levels of SPP1, S100A12, GAS6, and PROS1 at the time of hospital admission ( n = 92) stratified based on a patient’s subsequent need to be transferred to ICU. ( G ) Percentage of patients with acute COVID-19 pneumonia ( n = 92) transferred to ICU during the hospitalization based on having high levels of SPP1 (≥108 ng/mL), S100A12 (≥59 ng/mL), GAS6 (≥24 ng/mL), and PROS1 (≥15 μg/mL) at the time of hospital admission. ( B , D , and F ) Data are presented as violin plots with median and interquartile range. Asterisk indicates 1-way ANOVA (Kruskal-Wallis test) with Dunn’s correction for multiple comparisons if more than 2 groups were compared ( B ), or 2-sided Mann-Whitney U was used when 2 groups were compared ( B and D – G ). ( H ) Kaplan-Meier analysis of the rate of transfer of COVID-19 patients to ICU based on their cut-off values for SPP1, S100A12, GAS6, and PROS1 at the time of hospital admission.

Article Snippet: Blood samples were centrifuged (600 g /15 minutes) and plasma aliquots were stored at –80°C until analysis by ELISA for SPP1 (BMS2066; Thermo Fisher Scientific), S100A12 (DY1052; R&D Systems), GAS6 (DY885B; R&D Systems), and PROS1 (NBP2-60585; NOVUS Biological).

Techniques: Clinical Proteomics, MANN-WHITNEY

Journal: JCI Insight

Article Title: COVID-19 and RA share an SPP1 myeloid pathway that drives PD-L1 + neutrophils and CD14 + monocytes

doi: 10.1172/jci.insight.147413

Figure Lengend Snippet: ( A ) Representative images of lung CT scans (transversal and sagittal view) of a COVID-19 patient taken during acute pneumonia and during convalescence (68.60 ± 4.36 days after hospital discharge). ( B ) Plasma levels of SPP1, S100A12, GAS6, and PROS1 in paired plasma samples from COVID-19 patients at the time of acute pneumonia and at the convalescent phase ( n = 26). ( C ) Plasma levels of SPP1, S100A12, GAS6, and PROS1 in convalescent COVID-19 patients ( n = 41) stratified based on the severity of prior acute pneumonia and compared with the levels of healthy donors ( n = 10). ( D ) Plasma levels of IL-6 in acute pneumonias and post–COVID-19. ( E ) SPP1, S100A12, GAS6, and PROS1 in convalescent COVID-19 patients ( n = 41) stratified based on suffering ( n = 36) or not ( n = 5) at least 1 of the symptoms (fatigue, musculoskeletal, or respiratory symptoms). ( B ) Data are presented as before-and-after plot. Wilcoxon test on paired samples was used, and exact P values are provided on the graphs. ( C – E ) Data are presented as violin plots with median and interquartile range. Asterisks indicate 1-way ANOVA with correction for multiple comparisons if more than 2 groups were compared, or 2-sided Mann-Whitney U test was used when 2 groups were compared ( C – E ). Exact P values are provided on the graphs.

Article Snippet: Blood samples were centrifuged (600 g /15 minutes) and plasma aliquots were stored at –80°C until analysis by ELISA for SPP1 (BMS2066; Thermo Fisher Scientific), S100A12 (DY1052; R&D Systems), GAS6 (DY885B; R&D Systems), and PROS1 (NBP2-60585; NOVUS Biological).

Techniques: Clinical Proteomics, MANN-WHITNEY

Journal: Nature Communications

Article Title: Genome-wide DNA methylation profiling in blood reveals epigenetic signature of incident acute coronary syndrome

doi: 10.1038/s41467-024-51751-6

Figure Lengend Snippet: The 26 validated DMPs associated with incident ACS

Article Snippet: cg04517903 , 16:1557605 , TELO2 , 0.75 (0.67–0.84) , 0.72 (0.57–0.89) , 0.74 (0.67–0.82) , 8.36 ×10 −9.

Techniques:

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Prognostic Significance of PROS1 Downregulation in BLCA. ( A ) Gene intersections from DEGs of TCGA-BLCA cohort and two distinct subgroups and differential expressed proteins in proteomic datasets by Wan et al. ( B ) Comparative analysis of PROS1 mRNA levels in BLCA versus normal tissues in the GSE13507 cohort. ( C , D ) Assessment of PROS1 mRNA expression in BLCA compared to normal tissues within the TCGA cohort. ( E ) Pan-cancer analysis revealing widespread downregulation of PROS1 mRNA across various tumor types. ( F ) Correlation of clinicopathologic features with PROS1-defined subgroups. ( G ) Western blot analysis demonstrating PROS1 protein levels in eight paired BLCA and adjacent noncancerous tissues. ( H ) Immunohistochemical images showing PROS1 expression in BLCA and normal bladder tissues at 50× and 200× magnification. ( I , J ) Kaplan–Meier survival curves based on PROS1 IHC staining scores for OS and RFS. BLCA, bladder cancer. DEGs, differentially expressed genes. OS, overall survival. RFS, recurrence-free survival. * P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Immunohistochemistry

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Association between PROS1 expression clinicopathological factors of 45 patients in BLCA.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Univariate and Multivariate Cox regression models of 45 patients in BLCA.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques:

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Molecular Profiling of PROS1-Defined Subgroups. ( A ) Heatmap illustrating DEGs that are positively and negatively correlated with PROS1 expression. ( B ) GO enrichment analysis identifying biological processes (BP), cellular components (CC), and molecular functions (MF) associated with PROS1-defined subgroups. ( C ) KEGG pathway analysis highlighting enriched signaling pathways related to PROS1-defined subgroups. ( D ) DO enrichment analysis suggesting a correlated involvement of PROS1-defined subgroups in urinary bladder disease. DEGs, differentially expressed genes.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Expressing, Protein-Protein interactions

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Immune Characteristics of PROS1-Defined Subgroups. (A) Correlation analysis between PROS1-defined subgroups and TME scores. (B) Differential immune cell infiltration levels across PROS1-defined subgroups. (C) Association between PROS1-defined subgroups and various immune cell types. (D) Comparative analysis of immune checkpoint marker expression between subgroups. (E) Distribution of PROS1 expression among groups categorized by their clinical response to anti-PD-L1 therapy. TME, tumor microenvironment. * P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Marker, Expressing

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: PROS1 Overexpression Suppresses BLCA Cell Proliferation by Inducing Cell Cycle Arrest. (A, B) RT-qPCR and WB analyses demonstrating reduced PROS1 expression in multiple BLCA cell lines compared to normal urothelial cells. (C, D) Validation of PROS1 knockdown efficiency using shNC-PROS1, sh1-PROS1, sh2-PROS1, and sh3-PROS1 in UMUC-3 cells, and overexpression efficiency with NC-PROS1 and OE-PROS1 in J82 and T24 cell lines, confirmed by western blotting. (E-G) Assessment of cell proliferation through CCK8 and colony formation assays. (H, I) Flow cytometry analysis to examine cell cycle progression. (J) WB analysis of cell cycle-related regulatory proteins. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined by unpaired t-test or One-way ANOVA, followed by Dunnett’s multiple comparisons test (* P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001; ns indicates non-significant). RT-qPCR, real time quantitative polymerase chain reaction. WB, western blotting.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Biomarker Discovery, Knockdown, Western Blot, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: PROS1 overexpression inhibits migration, invasion, and angiogenesis in BLCA Cells. (A, B) Transwell assays evaluating cell migration and invasion capabilities. (C, D) WB analysis of EMT markers and MMPs. (E) Correlation between PROS1 and VEGFA mRNA expression levels based on TCGA data from TIMER 2. 0. (F, G) Analysis of VEGFA mRNA and protein expression in UMUC-3 sh2-PROS1, sh3-PROS1 cell lines, and J82 and T24 OE-PROS1 cell lines. (H) Evaluation of HUVEC tube formation using co-culture and Matrigel-based assays. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined by unpaired t-test or One-way ANOVA, followed by Dunnett’s multiple comparisons test (* P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001). BLCA, bladder cancer. WB, western blotting. EMT, epithelial-to-mesenchymal transition. MMP, matrix metalloproteinases. HUVEC, human umbilical vein endothelial cell.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Over Expression, Migration, Expressing, Co-Culture Assay, Western Blot

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: PROS1 Modulates BLCA Cell Phenotype via the AKT/GSK3β/β-Catenin Signaling Pathway. (A) Volcano plot of DEGs between NC-PROS1 and OE-PROS1 in J82 cell lines based on transcriptome sequencing data. (B) GO enrichment analysis identifying biological processes (BP), cellular components (CC), and molecular functions (MF) linked to DEGs between the groups. (C) KEGG pathway analysis highlighting signaling pathways enriched among DEGs between the groups. (D) Protein-protein interaction (PPI) network for DEGs derived from the STRING database. (E, F) WB analysis of AKT, p-AKT, GSK3β, p-GSK3β, and β-catenin expression in J82 and T24 cell lines following PROS1 overexpression. (G, H) Western blot analysis of AKT, p-AKT, GSK3β, p-GSK3β, and β-catenin expression in J82 and T24 cell lines after PROS1 overexpression and subsequent treatment with the AKT agonist SC79. BLCA, bladder cancer. DEGs, differentially expressed genes. WB, western blotting.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Sequencing, Protein-Protein interactions, Derivative Assay, Expressing, Over Expression, Western Blot

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Rescue experiments elucidating the role of PROS1 in BLCA Cell Behavior. (A-C) J82 and T24 cell lines transfected with OE-PROS1 and treated with the AKT agonist SC79 exhibited enhanced proliferative capacity in colony formation (A, B) and CCK8 assays (C) to some extent. (D) Flow cytometry confirmed SC79’s role in alleviating G1-S phase cell cycle arrest in both OE-PROS1 cell lines to a certain degree. (E, F) Transwell assays revealed increased migration and invasion capabilities in both OE-PROS1 cell lines following SC79 treatment. (G) Co-culture assays and Matrigel-based tube formation assays indicated that SC79 partially enhanced tube formation in HUVECs co-cultured with J82 and T24 OE-PROS1 cells. (H) WB analysis revealed a relative elevated VEGFA levels following SC79 treatment in J82 and T24 OE-PROS1 cells. Data are shown as mean ± SD from three independent experiments. Statistical significance was determined by one-way or two-way ANOVA, followed by Tukey’s multiple comparisons test (* P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001). BLCA, bladder cancer. HUVEC, human umbilical vein endothelial cells. WB, western blotting.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Transfection, Flow Cytometry, Migration, Co-Culture Assay, Cell Culture, Western Blot

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: In vivo inhibition of tumorigenicity by PROS1 overexpression. ( A , B ) Tumor size and volume were measured at 7-day intervals over 28 days post-injection to construct tumor growth curves. ( C ) Tumor weight was measured on day 28 post-injection in nude mice from each group. ( D ) H&E staining was employed to assess BLCA cell morphology. ( E ) IHC staining confirmed ectopic PROS1 expression; Ki-67 staining demonstrated a significant reduction in cell proliferation, while VEGFA staining revealed decreased VEGFA protein levels in J82 xenografts expressing PROS1. Scale bar = 50 μm. Data are shown as mean ± SD from from six mice per group. Statistical significance was determined by two-way ANOVA, followed by Šídák’s multiple comparisons test or unpaired t-test (* P < 0. 05, ** P < 0. 01, *** P < 0. 001, **** P < 0. 0001). H&E, hematoxylin and eosin. BLCA, bladder cancer. IHC, immunohistochemistry.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: In Vivo, Inhibition, Over Expression, Injection, Construct, Staining, Immunohistochemistry, Expressing

Journal: Scientific Reports

Article Title: Mechanistic insights into PROS1 inhibition of bladder cancer progression and angiogenesis via the AKT/GSK3β/β-catenin pathway

doi: 10.1038/s41598-025-89217-4

Figure Lengend Snippet: Sequences of primers for RT-qPCR.

Article Snippet: After a 20-minute blocking step with protein-free rapid blocking buffer (Servicebio, #G2052), the membrane was incubated overnight at 4 °C with the primary antibody, including GAPDH (Proteintech, #10494-1-AP), β-Actin (Proteintech, #66009-1-Ig), PROS1 (Abmart, #T58808), PROS1 (Proteintech, #16910-1-AP), AKT (Proteintech, #10176-2-AP), p-AKT (Proteintech, #66444-1-Ig), GSK3β (Proteintech, #22104-1-AP), p-GSK3β (Proteintech, #14850-1-AP), β-catenin (Proteintech, #51067-2-AP), VEGFA (Proteintech, #19003-1-AP), CDK4 (Proteintech, #11026-1-AP), CDK6 (Proteintech, #14052-1-AP), CyclinD1 (Proteintech, #60186-1-Ig), Vimentin (Proteintech, #22031-1-AP), MMP-9 (Proteintech, # 30592-1-AP), N-cadherin (Proteintech, #22018-1-AP), E-cadherin (Proteintech, #60186-1-Ig), Ki67 (Proteintech, #27309-1-AP), p27 (MCE, #HY-P80256), and PCNA (MCE, #HY-P80268).

Techniques: Sequencing