primary antibodies against survivin  (Novus Biologicals)

Journal: bioRxiv

Article Title: Survivin as a Multifaceted Oncogenic Driver and Therapeutic Target in Renal Cell Carcinoma

doi: 10.1101/2025.09.02.673805

Figure Lengend Snippet: A) Immunoblot of survivin expression in control vs survivin siRNA treated RENCA cells. Tubulin serves as loading control. B) Bar graph depicting relative cell proliferation between ctrl vs survivin siRNA treated RENCA cells. Proliferation was assessed via cell counting. C-E) G0-synchronized RENCA cells were treated with 0.5μM, 1μM, and 2μM YM155 for 24 hours. Whole cell lysates were collected and analyzed with immunoblotting (C) to determine protein levels of Survivin (D) and Cyclin D1 (E). Expression levels were normalized to α-tubulin and displayed as a fold change to DMSO. n=4. G0-synchronized RENCA cells were treated with 1μM and 2μM YM155 for 24 hours. F-G) Expression levels were normalized to GAPDH and displayed as a fold change to DMSO. n=4. S-phase entry with YM155 treatment (F) and adenoviral overexpression (G) was assessed by EdU incorporation. Percentage of EdU positive cells was normalized to values from RENCA cells treated with DMSO (F) or GFP adenovirus (G). n=4. Data are means + SEMs. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: Primary antibodies against survivin (NB500-201, Novus Biologicals; 1:500) and cyclin D1 (sc-20044, Santa Cruz Biotechnology; 1:200), were incubated at room temperature for 1 hour and overnight at 4°C.

Techniques: Western Blot, Expressing, Control, Cell Counting, Over Expression

Journal: APL Bioengineering

Article Title: Survivin modulates stiffness-induced vascular smooth muscle cell motility

doi: 10.1063/5.0252766

Figure Lengend Snippet: Survivin knockdown decreases stiffness-stimulated cell migration. (a) VSMCs transfected with 200 nM survivin siRNA or non-targeting control siRNA were seeded on fibronectin-coated stiff hydrogels for 4 h. (b) Total cell lysates were analyzed by immunoblotting, and survivin protein levels were normalized to GAPDH. n = 3 independent biological replicates. (c)–(e) VSMCs were plated for time-lapse video microscopy, and manual tracking of these cells was conducted to obtain single-cell trajectories (c) and average cell velocities (d). 37 cells (control siRNA) and 37 cells (survivin siRNA) were analyzed from n = 3 independent biological replicates. (e) Sequences of images from a set of representative time-lapse experiments (arrow-direction of protrusion). * p < 0.05, *** p < 0.001.

Article Snippet: The PVDF membrane was blocked in 5% milk in TBST (Tris-buffered saline with 0.1% Tween 20 detergent) for 1 h at room temperature, then incubated with primary antibodies against survivin (1:250; Cat. No. 71G4B7, Cell Signaling Technology) and GAPDH (1:10 000; Cat. No. 60004-1-Ig, Proteintech) diluted in 5% milk in TBST for 2 h at room temperature, overnight at 4 °C, and an additional 2 h at room temperature followed by a 30-min wash with TBST.

Techniques: Knockdown, Migration, Transfection, Control, Western Blot, Microscopy

Journal: APL Bioengineering

Article Title: Survivin modulates stiffness-induced vascular smooth muscle cell motility

doi: 10.1063/5.0252766

Figure Lengend Snippet: Survivin regulates FAK phosphorylation, actin organization, and stress fiber formation. (a) VMSCs were sparsely seeded on fibronectin-coated soft and stiff hydrogels and treated with either YM155 or DMSO. Cells were stained with DAPI, phospho-FAK (Tyr397) antibody and Alexa Fluor 647-phalloidin. Fluorescence images were acquired using a spinning disk confocal microscope with a 63× oil-immersion objective. The outlined boxes indicate the regions shown in the magnified insets. (b) A sum of slices projection of each cell was generated using Fiji/ImageJ, and FAK pY397 clusters were manually counted. 37 cells (soft-DMSO), 44 cells (stiff-DMSO), 33 cells (stiff-0.1 µM YM155), and 37 cells (stiff-2µM YM155) were analyzed from n = 3 independent biological replicates. * p < 0.05, ** p < 0.01.

Article Snippet: The PVDF membrane was blocked in 5% milk in TBST (Tris-buffered saline with 0.1% Tween 20 detergent) for 1 h at room temperature, then incubated with primary antibodies against survivin (1:250; Cat. No. 71G4B7, Cell Signaling Technology) and GAPDH (1:10 000; Cat. No. 60004-1-Ig, Proteintech) diluted in 5% milk in TBST for 2 h at room temperature, overnight at 4 °C, and an additional 2 h at room temperature followed by a 30-min wash with TBST.

Techniques: Phospho-proteomics, Staining, Fluorescence, Microscopy, Generated

Journal: APL Bioengineering

Article Title: Survivin modulates stiffness-induced vascular smooth muscle cell motility

doi: 10.1063/5.0252766

Figure Lengend Snippet: Survivin overexpression partially rescues cell migration on soft hydrogels. (a) VSMCs were infected with either adenoviral GFP (50 MOI) or survivin (25 and 50 MOI). (b) Total cell lysates were analyzed by immunoblotting, and survivin protein levels were normalized to GAPDH. n = 3 independent biological replicates. (c)–(e) VSMCs were plated for time-lapse video microscopy, and manual tracking of these cells was conducted to obtain single-cell trajectories (c) and average cell velocities (d). 59 cells (GFP 50 MOI), 60 cells (survivin 25 MOI), and 60 cells (survivin 50 MOI) were analyzed from n = 5 independent biological replicates. (e) Sequences of images from a set of representative time-lapse experiments (arrow-direction of protrusion). * p < 0.05, *** p < 0.001 . ns, not significant.

Article Snippet: The PVDF membrane was blocked in 5% milk in TBST (Tris-buffered saline with 0.1% Tween 20 detergent) for 1 h at room temperature, then incubated with primary antibodies against survivin (1:250; Cat. No. 71G4B7, Cell Signaling Technology) and GAPDH (1:10 000; Cat. No. 60004-1-Ig, Proteintech) diluted in 5% milk in TBST for 2 h at room temperature, overnight at 4 °C, and an additional 2 h at room temperature followed by a 30-min wash with TBST.

Techniques: Over Expression, Migration, Infection, Western Blot, Microscopy

Journal: Discover Oncology

Article Title: Increased protein expression of interleukin-10 and its signalling molecules in colon cancer progression: potential prognostic and therapeutic targets

doi: 10.1007/s12672-025-02452-z

Figure Lengend Snippet: Protein detection of total and phosphorylated STAT3 by Western blotting alongside their relative expression (mean ± SD) in non-treated control cells (CT) and after treatment with varying concentrations (0.1, 1, and 40 ng/ml) of IL-10 protein and anti-IL-10Rα monoclonal antibody, and a single concentration of STAT3 inhibitor V (2 µM) for 24 h in the ( a ) HT29 and ( b ) SW620 human colon cancer cell lines (mean ± SD; a = P < 0.01 compared to the CT group; b = P < 0.01 compared to 0.1 ng/mL of IL-10 protein; c = P < 0.01 compared to 1 ng/mL of IL-10 protein; d = P < 0.01 compared to 40 ng/mL of IL-10 protein; e = P < 0.01 compared to 0.1 ng/mL of anti-IL-10Rα antibody; f = P < 0.01 compared to 1 ng/mL of anti-IL-10Rα antibody; and g = P < 0.01 compared to 40 ng/mL of anti-IL-10Rα antibody). The uncropped Western blot membranes for total and phosphorylated STAT3 are provided in Suppl. Figure 4

Article Snippet: Western blot analysis was performed to assess protein expression levels of CCND1 (#2978), CCND3 (#2936), p21 (#37543), p27 (#3686), PCNA (#13110), survivin (#2808), and cleaved Caspase-3 (#9664) using primary rabbit monoclonal antibodies from Cell Signaling Technology Inc. (MA, USA).

Techniques: Western Blot, Expressing, Control, Concentration Assay

Journal: Discover Oncology

Article Title: Increased protein expression of interleukin-10 and its signalling molecules in colon cancer progression: potential prognostic and therapeutic targets

doi: 10.1007/s12672-025-02452-z

Figure Lengend Snippet: Distribution of cells (mean ± SD) in the different phases of cell cycle in non-treated control cells (CT) and after treatment with varying concentrations (0.1, 1, and 40 ng/ml) of IL-10 protein and anti-IL-10Rα monoclonal antibody, or a single concentration of STAT3 inhibitor (2 µM) for 24 h in the ( a ) HT29 and ( b ) SW620 human colon cancer cell lines (mean ± SD; a = P < 0.01 compared to the CT group; b = P < 0.01 compared to 0.1 ng/mL of IL-10 protein; c = P < 0.01 compared to 1 ng/mL of IL-10 protein; d = P < 0.01 compared to 40 ng/mL of IL-10 protein; e = P < 0.01 compared to 0.1 ng/mL of anti-IL-10Rα antibody; f = P < 0.01 compared to 1 ng/mL of anti-IL-10Rα antibody; and g = P < 0.01 compared to 40 ng/mL of anti-IL-10Rα antibody)

Article Snippet: Western blot analysis was performed to assess protein expression levels of CCND1 (#2978), CCND3 (#2936), p21 (#37543), p27 (#3686), PCNA (#13110), survivin (#2808), and cleaved Caspase-3 (#9664) using primary rabbit monoclonal antibodies from Cell Signaling Technology Inc. (MA, USA).

Techniques: Control, Concentration Assay

Journal: Discover Oncology

Article Title: Increased protein expression of interleukin-10 and its signalling molecules in colon cancer progression: potential prognostic and therapeutic targets

doi: 10.1007/s12672-025-02452-z

Figure Lengend Snippet: a Heatmap showing relative mRNA expression (mean ± SD) of CCND1, CCND3, p21, and p27 genes alongside ( b ) their relative protein expression (mean ± SD) by Western blotting, in non-treated control cells (CT) and after treatment with varying concentrations (0.1, 1, and 40 ng/ml) of IL-10 protein and anti-IL-10Rα monoclonal antibody, or a single concentration of STAT3 inhibitor V (2 µM) for 24 h in the HT29 and SW620 human colon cancer cell lines (mean ± SD; a = P < 0.01 compared to the CT group; b = P < 0.01 compared to 0.1 ng/mL of IL-10 protein; c = P < 0.01 compared to 1 ng/mL of IL-10 protein; d = P < 0.01 compared to 40 ng/mL of IL-10 protein; e = P < 0.01 compared to 0.1 ng/mL of anti-IL-10Rα antibody; f = P < 0.01 compared to 1 ng/mL of anti-IL-10Rα antibody; and g = P < 0.01 compared to 40 ng/mL of anti-IL-10Rα antibody). The uncropped Western blot membranes for CCND1, CCND3, p21, and p27 proteins are provided in Suppl. Figure 4

Article Snippet: Western blot analysis was performed to assess protein expression levels of CCND1 (#2978), CCND3 (#2936), p21 (#37543), p27 (#3686), PCNA (#13110), survivin (#2808), and cleaved Caspase-3 (#9664) using primary rabbit monoclonal antibodies from Cell Signaling Technology Inc. (MA, USA).

Techniques: Expressing, Western Blot, Control, Concentration Assay

Journal: Discover Oncology

Article Title: Increased protein expression of interleukin-10 and its signalling molecules in colon cancer progression: potential prognostic and therapeutic targets

doi: 10.1007/s12672-025-02452-z

Figure Lengend Snippet: Percentage (mean ± SD) of living, early and late apoptotic, and dead cells in non-treated control cells (CT) and after treatment with varying concentrations (0.1, 1, and 40 ng/ml) of IL-10 protein and anti-IL-10Rα monoclonal antibody, or a single concentration of STAT3 inhibitor V (2 µM) for 24 h in the ( a ) HT29 and ( b ) SW620 human colon cancer cell lines (mean ± SD; a = P < 0.01 compared to the CT group; b = P < 0.01 compared to 0.1 ng/mL of IL-10 protein; c = P < 0.01 compared to 1 ng/mL of IL-10 protein; d = P < 0.01 compared to 40 ng/mL of IL-10 protein; e = P < 0.01 compared to 0.1 ng/mL of anti-IL-10Rα antibody; f = P < 0.01 compared to 1 ng/mL of anti-IL-10Rα antibody; and g = P < 0.01 compared to 40 ng/mL of anti-IL-10Rα antibody)

Article Snippet: Western blot analysis was performed to assess protein expression levels of CCND1 (#2978), CCND3 (#2936), p21 (#37543), p27 (#3686), PCNA (#13110), survivin (#2808), and cleaved Caspase-3 (#9664) using primary rabbit monoclonal antibodies from Cell Signaling Technology Inc. (MA, USA).

Techniques: Control, Concentration Assay

Journal: Discover Oncology

Article Title: Increased protein expression of interleukin-10 and its signalling molecules in colon cancer progression: potential prognostic and therapeutic targets

doi: 10.1007/s12672-025-02452-z

Figure Lengend Snippet: a Heatmap showing relative mRNA expression (mean ± SD) of PCNA, survivin, and caspase-3 genes alongside ( b ) their relative protein expression (mean ± SD) by Western blotting, in non-treated control cells (CT) and after treatment with varying concentrations (0.1, 1, and 40 ng/ml) of IL-10 protein and anti-IL-10Rα monoclonal antibody, or a single concentration of STAT3 inhibitor V (2 µM) for 24 h in the HT29 and SW620 human colon cancer cell lines (mean ± SD; a = P < 0.01 compared to the CT group; b = P < 0.01 compared to 0.1 ng/mL of IL-10 protein; c = P < 0.01 compared to 1 ng/mL of IL-10 protein; d = P < 0.01 compared to 40 ng/mL of IL-10 protein; e = P < 0.01 compared to 0.1 ng/mL of anti-IL-10Rα antibody; f = P < 0.01 compared to 1 ng/mL of anti-IL-10Rα antibody; g = P < 0.01 compared to 40 ng/mL of anti-IL-10Rα antibody). The uncropped Western blot membranes for PCNA, survivin, and caspase-3 proteins are provided in Suppl. Figure 4

Article Snippet: Western blot analysis was performed to assess protein expression levels of CCND1 (#2978), CCND3 (#2936), p21 (#37543), p27 (#3686), PCNA (#13110), survivin (#2808), and cleaved Caspase-3 (#9664) using primary rabbit monoclonal antibodies from Cell Signaling Technology Inc. (MA, USA).

Techniques: Expressing, Western Blot, Control, Concentration Assay