primary antibodies against sphk1  (Novus Biologicals)

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 3. Correlation of SPHK1 gene expression with various clinicopathological characteristics in breast tumor tissues. (A) pN Stage (pN0 + pNx- [Nodal Negative], pN1 + pN2-[Nodal Positive]) (B) pTNM Stage (pTNM I + II- [early stage], pTNM III + IV- [late stage]) (C) Correlation with Ki67.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Gene Expression

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 4. Protein expression of SPHK1 and CERK in breast cancer patients. (A) Representative blots in adjacent normal (N) and tumor (T) tissues, (B) Densitometric analysis of SPHK1 and (C) CERK levels in adjacent normal and tumor tissues.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Expressing

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 6. Network-based analysis of interaction network corresponding to candidate genes. (A) The node degree distribution of PPI network. The number of genes is plotted as a function of their degree reflecting a power-law like distribution. The red line corresponds to a power-law distribution. (B) Sub-network of candidate genes. The large-sized black-colored nodes represent the candidate genes (ABCC1, ABCG2, CERK, MMP-2, MMP-9, and SPHK1), while the small gray color nodes represent the corresponding interactor genes. Interactions are represented in gray color and the depth of color represents the strength of correlations. (C) Shortest path lengths among candidate genes. Heatmap of shortest path length among the candidate genes, where the values represent the number of shortest paths between any pair.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 8. Correlation of CERK with (A) MMP-2, (B) MMP-9 and SPHK1 with (C) MMP-2, (D) MMP-9 in local cohort and CERK with (E) MMP-2, (F) MMP-9 and SPHK1 with (G) MMP-2, (H) MMP-9 in TCGA cohort.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 10. Correlation of CERK with (A) ABCC1, (B) ABCG2 and SPHK1 with (C) ABCC1, (D) ABCG2 in local cohort and CERK with (E) ABCC1, (F) ABCG2 and SPHK1 with (G) ABCC1, (H) ABCG2 in TCGA cohort.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 11. Expression of SPNS2 in TCGA cohort. (A) Relative gene expression between adjacent normal and tumor tissue, (B) Correlation with SPHK1.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Expressing, Gene Expression

Journal: Frontiers in Molecular Biosciences

Article Title: Neuroprotective Effects of Danshen Chuanxiongqin Injection Against Ischemic Stroke: Metabolomic Insights by UHPLC-Q-Orbitrap HRMS Analysis

doi: 10.3389/fmolb.2021.630291

Figure Lengend Snippet: (A,B) Western blot analysis of Sphk1 and S1PR1 in the cerebral cortex and hippocampus of rats at 24h after MCAO. (C,D) Representative photographs represented the expression of CD62P in the cerebral cortex and hippocampus at 24h reperfusion after MCAO. The positive products were brown granules after immunostaining, which was remarkably increased followed MCAO and was significantly decreased after treatment with DSCXQ. Representative images of Sphk1 and S1PR1 were presented, and the protein levels were expressed as a ratio of the β-tubulin levels ( n = 3). Data were expressed with mean values ± standard deviation (SD). ## p < 0.01 vs. Sham group, ** p < 0.01 * p < 0.05 vs. Model group.

Article Snippet: Membranes were probed with primary antibodies against Sphk1 (1:1000), S1PR1 (1:1000) (Affinity Biosciences, OH, United States), Bcl-2, Bax (1:1000), Cleaved Caspase-3 (1:500) (Cell Signaling Technology, Boston, MA, United States) at 4°C overnight.

Techniques: Western Blot, Expressing, Immunostaining, Standard Deviation

Journal: Pulmonary Circulation

Article Title: SphK1/S1P mediates TGF-β1-induced proliferation of pulmonary artery smooth muscle cells and its potential mechanisms

doi: 10.1177/2045894018816977

Figure Lengend Snippet: (a) PASMC were treated with different concentration of TGF-β1 in the range of 0–30 ng/mL for 24 h; the protein level of SphK1 was detected using western blotting, β-actin served as a loading control (n = 5 each group). (b) PASMC were exposed to 10 ng/mL TGF-β1 for the indicated times; the protein level of SphK1 was detected using western blotting, β-actin served as a loading control (n = 5 each group). * P < 0.05 vs. control.

Article Snippet: After blocking for 1 h at room temperature with 5% non-fat milk, the membranes were incubated overnight at 4°C with primary antibodies against SphK1 (1:1000 dilution; Cell Signaling Technology, USA), p-Smad2/3 (1:800 dilution; Cell Signaling Technology, USA), t-Smad2/3 (1:1000 dilution; Abcam, Cambridge, UK), NICD3 (1:500 dilution; Abcam, Cambridge, UK), and β-actin (1:1000 dilution; Santa Cruz, USA).

Techniques: Concentration Assay, Western Blot, Control

Journal: Pulmonary Circulation

Article Title: SphK1/S1P mediates TGF-β1-induced proliferation of pulmonary artery smooth muscle cells and its potential mechanisms

doi: 10.1177/2045894018816977

Figure Lengend Snippet: (a) PASMC were first incubated with or without 10 μM SB431542 for 1 h before stimulation with 10 ng/mL TGF-β1 for 1 h; the protein levels of p-Smad2/3 and t-Smad2/3 were detected using western blotting (n = 5 each group). (b) PASMC were first incubated with or without 10 μM SB431542 for 1 h before stimulation with 10 ng/mL TGF-β1 for 24 h; the protein level of SphK1 was detected using western blotting; β-actin served as a loading control (n = 5 each group). (c) PASMC were first incubated with or without 10 μM SB431542 for 1 h before stimulation with 10 ng/mL TGF-β1 for 24 h; levels of S1P in cell lysates and cell culture supernates were determined using S1P ELISA kits (n = 5 each group). * P < 0.05 vs. control, # P < 0.05 vs. TGF-β1-treated cells.

Article Snippet: After blocking for 1 h at room temperature with 5% non-fat milk, the membranes were incubated overnight at 4°C with primary antibodies against SphK1 (1:1000 dilution; Cell Signaling Technology, USA), p-Smad2/3 (1:800 dilution; Cell Signaling Technology, USA), t-Smad2/3 (1:1000 dilution; Abcam, Cambridge, UK), NICD3 (1:500 dilution; Abcam, Cambridge, UK), and β-actin (1:1000 dilution; Santa Cruz, USA).

Techniques: Incubation, Western Blot, Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Pulmonary Circulation

Article Title: SphK1/S1P mediates TGF-β1-induced proliferation of pulmonary artery smooth muscle cells and its potential mechanisms

doi: 10.1177/2045894018816977

Figure Lengend Snippet: (a) PASMC were transfected with SphK1 sequence-specific siRNA and non-targeting siRNA; SphK1 protein level was determined using western blotting; β-actin served as a loading control (n = 5 each group). (b) PASMC were first transfected with SphK1-specific or non-targeting siRNA for 24 h and then treated with 10 ng/mL TGF-β1 for 24 h; protein level of NICD3 was detected using western blotting, β-actin served as a loading control (n = 5 each group). * P < 0.05 vs. control, # P < 0.05 vs. TGF-β1-treated cells.

Article Snippet: After blocking for 1 h at room temperature with 5% non-fat milk, the membranes were incubated overnight at 4°C with primary antibodies against SphK1 (1:1000 dilution; Cell Signaling Technology, USA), p-Smad2/3 (1:800 dilution; Cell Signaling Technology, USA), t-Smad2/3 (1:1000 dilution; Abcam, Cambridge, UK), NICD3 (1:500 dilution; Abcam, Cambridge, UK), and β-actin (1:1000 dilution; Santa Cruz, USA).

Techniques: Transfection, Sequencing, Western Blot, Control

Journal: Pulmonary Circulation

Article Title: SphK1/S1P mediates TGF-β1-induced proliferation of pulmonary artery smooth muscle cells and its potential mechanisms

doi: 10.1177/2045894018816977

Figure Lengend Snippet: PASMC were pre-incubated with 10 μM SB431542 for 1 h or prior transfected with SphK1 siRNA for 24 h or pre-incubated with 10 μM DAPT for 4 h and then stimulated with 10 ng/mL TGF-β1 for an additional 24 h; the proliferation of cells was evaluated by MTT (n = 6 each group). * P < 0.05 vs. control, # P < 0.05 vs. TGF-β1-treated cells, & P < 0.05 vs. control siRNA + TGF-β1-treated group.

Article Snippet: After blocking for 1 h at room temperature with 5% non-fat milk, the membranes were incubated overnight at 4°C with primary antibodies against SphK1 (1:1000 dilution; Cell Signaling Technology, USA), p-Smad2/3 (1:800 dilution; Cell Signaling Technology, USA), t-Smad2/3 (1:1000 dilution; Abcam, Cambridge, UK), NICD3 (1:500 dilution; Abcam, Cambridge, UK), and β-actin (1:1000 dilution; Santa Cruz, USA).

Techniques: Incubation, Transfection, Control

Journal: Pulmonary Circulation

Article Title: SphK1/S1P mediates TGF-β1-induced proliferation of pulmonary artery smooth muscle cells and its potential mechanisms

doi: 10.1177/2045894018816977

Figure Lengend Snippet: TGF-β1 upregulates SphK1 expression and S1P production by promoting phosphorylation of Smad2/3 in primary cultured PASMC; SphK1/S1P then further mediates TGF-β1-induced Notch3 activation and ultimately mediates TGF-β1-induced PASMC proliferation.

Article Snippet: After blocking for 1 h at room temperature with 5% non-fat milk, the membranes were incubated overnight at 4°C with primary antibodies against SphK1 (1:1000 dilution; Cell Signaling Technology, USA), p-Smad2/3 (1:800 dilution; Cell Signaling Technology, USA), t-Smad2/3 (1:1000 dilution; Abcam, Cambridge, UK), NICD3 (1:500 dilution; Abcam, Cambridge, UK), and β-actin (1:1000 dilution; Santa Cruz, USA).

Techniques: Expressing, Phospho-proteomics, Cell Culture, Activation Assay