Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) Relative SHP-1 protein expression in the 19 BCa cell lines examined in this study. ( B ) Expression levels were significantly higher in epithelial-like cells (n = 11) than in intermediate and mesenchymal-like lines (n = 8). Error bars represent the standard error of the mean (SEM).

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) SHP-1 protein expression in bladder tissue was significantly lower in muscle-invasive (MI, n = 8) tumor than in non-muscle-invasive (NMI, n = 8) and urothelial tissue with no residual disease (NRD, n = 10). ( B ) Examples of DAB and corresponding H&E staining in the bladder tissues examined in this study. The error bars represent the interquartile range.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Staining

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Correlation plots (n = 19) of SHP-1 protein expression with the EMT markers E-cadherin, N-cadherin, and Vimentin. Gray areas represent the 95%CI. Representative Western blot images: from the BCa cell lines EJ and HT-1376, the first lane of each pair corresponds to the protein indicated above each lane, and the second corresponds to the total protein detected for the same lane.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Western Blot

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) SHP-1 protein expression following transduction with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 6, RT-112 n = 7, TCCSUP n = 4, UM-UC-3 n = 6. ( B ) Representative Western blot images: the first lane of each pair corresponds to SHP-1, and the second corresponds to the total protein detected for the same lane. Error bars represent the SEM. ( C ) Relative proliferation rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 5, RT-112 n = 5, TCCSUP n = 9, UM-UC-3 n = 5. Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Transduction, shRNA, Western Blot

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) Relative migration rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 5, RT-112 n = 4, TCCSUP n = 3, UM-UC-3 n = 3. ( B ) Representative images from in vitro migration assays (scale bar = 200 μm). ( C ) Relative invasion rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 3, RT-112 n = 5, TCCSUP n = 3, UM-UC-3 n = 3. ( D ) Representative images from in vitro invasion assays (scale bar = 200 μm). Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Migration, Transduction, shRNA, In Vitro

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Significant gene sets resulting from GSEA of RNA sequence data comparing the transduced BCa lines with high-SHP-1-expressing lines versus low-SHP-1-expressing lines (q-value < 0.1).

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Sequencing, Expressing

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Relative expression of pAkt/Akt in BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): RT-112 n = 7 and TCCSUP n = 8. Representative Western blot images: the first lane of each pair corresponds to Akt or pAkt (ser473), and the second corresponds to the total protein detected for the same lane. Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Transduction, shRNA, Western Blot

Journal: Nature chemical biology

Article Title: A druggable redox switch on SHP1 controls macrophage inflammation

doi: 10.1038/s41589-026-02163-8

Figure Lengend Snippet: a, LPS-induced (100 ng/ml) SHP1 Tyr536, Tyr564 and Ser591 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. b , LPS-induced (100 ng/ml) STAT3 Tyr705 and Ser727 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. c,e,f , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs pre-incubated 3 h with SCA1, SCA1-NC or the respective derivatives over a dose response (0.313–80 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 4). For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, iBMDMs were pre-treated with LPS (100 ng/ml) for 3 h followed by treatment with DMSO or SCA9 at the indicated concentrations for 45 min and primed with adenosine triphosphate (ATP, 5 mM) for 45 min ( n = 3). Absolute cytokine production as a function of concentration is used to calculate IC 50 values. d , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs treated 3 h with SCA1 (20 μM) alone or 6 h with LPS (100 ng/ml) alone ( n = 3). g, Phagocytic activity of iBMDMs pre-treated 3 h with SCA1, SCA9, SCA7 or SCA25 (10 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 6). h, Relative fold changes in protein abundance between iBMDMs treated with SCA1 (50 μM) for 3 h followed by LPS (100 ng/ml, 15 min) versus LPS alone ( n = 3). Data are mean ± s.e.m. (in d,h ) or s.d. (in c,e-g ). P values calculated using one-way or two-way ANOVA for multiple comparisons or two-tailed Student’s t -tests for unpaired comparisons.

Article Snippet: Equal amounts of proteins were resolved using NuPAGE 4–12% Bis-Tris 15-well or 26-well gels (Invitrogen, #NP0336BOX or #WG1403BOX), transferred onto PVDF membranes (Thermo Fisher Scientific, iBlot 2 Gel Transfer Device, #IB21001), and immunoblotted with anti-IRAK1 (Cell Signaling, #4504, rabbit mAb; 1:1000 dilution), anti-phospho-IκBα (Ser32/36) (Cell Signaling, #9246; mouse mAb; 1:1000 dilution), anti-IκBα (Cell Signaling, #9242, rabbit pAb; 1:1000 dilution), anti-phospho-NFκB p65 (Ser536) (Cell Signaling, #3033, rabbit mAb; 1:1000 dilution), anti-NFκB p65 (Cell Signaling, #8242, rabbit mAb; 1:1000 dilution), anti-SHP1 (Cell Signaling, #3759, rabbit mAb; 1:1000 dilution), anti-phospho-SHP1 (Tyr536) (Invitrogen, #PA5–36682, rabbit pAb; 1:1000 dilution), anti-phospho-SHP1 (Tyr564) (Cell Signaling, #8849, rabbit mAb; 1:1000 dilution), anti-phospho-SHP1 (Ser591) (Invitrogen, #PA5–143764, rabbit pAb; 1:1000 dilution), anti-STAT3 (Cell Signaling, #4904, rabbit mAb; 1:1000 dilution), anti-phospho-STAT3 (Tyr705) (Cell Signaling, #9145, rabbit mAb; 1:1000 dilution), anti-phospho-STAT3 (Ser727) (Cell Signaling, #9136, mouse mAb; 1:1000 dilution) and anti-β-actin (Cell Signaling, #3700, mouse mAb; 1:3000 dilution) antibodies, diluted in TBS-T (Boston BioProducts, Inc., #IBB-180X) containing 5% BSA, followed by incubation with horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibodies (Promega, #W4011 or #W4021), diluted 1:10000 in TBS-T containing 5% milk, and visualized using ECL western blotting substrates (Pierce ECL Western Blotting Substrate, #32106).

Techniques: Phospho-proteomics, Western Blot, Incubation, Concentration Assay, Activity Assay, Quantitative Proteomics, Two Tailed Test