primary antibodies against actb (actin beta)  (ABclonal Biotechnology)

Journal: Cancer letters

Article Title: SIRPA enhances osteosarcoma metastasis by stabilizing SP1 and promoting SLC7A3-mediated arginine uptake.

doi: 10.1016/j.canlet.2023.216412

Figure Lengend Snippet: Fig. 7. SP1 regulates SLC7A3 expression. A-B. mRNA expression (A) and protein level (B) of SLC7A3 in OS cells with SP1 knockdown or overexpression (n = 3). C. Arginine uptake ratio in OS cells with/without SP1 knockdown upon SLC7A3 overexpression (n = 3). D. Schematic diagram showing possible regulatory sites for SP1 in the SLC7A3 promoter predicted by the JASPAR database (top). Construction of the SLC7A3 luciferase plasmids: wild-type SLC7A3 promoter (WT) and deletion mutants lacking region 1 (ΔRegion 1), region 2 (ΔRegion 2), or both (ΔRegion 1 + 2) (bottom). E. Homology analysis of region 1 and region 2 in the SLC7A3 promoter. F. 143B cells with/without SIRPA knockdown were transfected with plasmids encoding SP1(-500/0)-Luc WT, ΔRegion1, ΔRegion2, and ΔRegion1+2. Then, luciferase levels were measured in extracts after 48 h. G-H. ChIP assay of DNA fragments containing putative region 1 and region 2 within the SLC7A3 promoter region followed by PCR. The SLC7A3 promotor region beyond SP1 regulation (CTR) and the β-actin region (ACTB) were used as negative controls; an anti- histone antibody and rabbit IgG were used as assay controls (G). The histogram shows the relative amount of each DNA fragment binding to SP1 (H). The data are presented as the means±SDs. *P < 0.05, **P < 0.01 by one-way ANOVA.

Article Snippet: The membranes were then incubated with diluted (1:1000) primary antibodies against human ACTB (4970; Cell Signaling Technology), SIRPA (47027; Cell Signaling Technology), SLC7A3 (PA5-103721; Invitrogen), RNF4 (12187; Cell Signaling Technology), ubiquitin (3936; Cell Signaling Technology), SP1 (9389; Cell Signaling Technology), p-Thr278-SP1 (PA5-106039; Invitrogen), p-Thr739-SP1 (PA5-104771; Invitrogen), ERK (4695; Cell Signaling Technology), p-ERK (9101; Cell Signaling Technology), AKT (4691; Cell Signaling Technology), p-AKT (4060; Cell Signaling Technology), β-catenin (8480; Cell Signaling Technology), and p-β-catenin (9567; Cell Signaling Technology).

Techniques: Expressing, Knockdown, Over Expression, Luciferase, Transfection, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Pentagalloyl Glucose, a Major Compound in Mango Seed Kernel, Exhibits Distinct Gastroprotective Effects in Indomethacin-Induced Gastropathy in Rats via Modulating the NO/eNOS/iNOS Signaling Pathway

doi: 10.3389/fphar.2022.800986

Figure Lengend Snippet: Upper panel represents the effect of pretreatment with either pentagalloyl glucose (PGG) (100 or 200 mg/kg) or famotidine on gastric PECAM-1 level of indomethacin-induced gastric injury. The lower panel represents densitometry analysis of the ratio of PECAM-1 protein over β-actin protein. Data are expressed as mean ± SEM, n = 3. *, @ Statistically significant from control and indomethacin groups of rats, respectively, at p < 0.05 by one-way ANOVA was used for statistical analysis followed by Tukey’s post hoc test.

Article Snippet: Ten percent SDS-PAGE was used to separate tissue proteins, which were then transferred to nitrocellulose membranes using the Trans-Blot ® semi-dry transfer cell (Bio- Rad, Hercules, CA, United States) at 20 v for 15 min. After blocking with 1 × Tris-buffered saline/0.1% Tween 20 (TBST) with 5% non-fat dry milk for 1 h, rabbit polyclonal antibodies against VEGF (Catalog no: CAB12303; Assay genie, Dublin, Ireland; dilution, 1:500); primary rabbit polyclonal antibodies against PECAM-1 (Catalog no: PA5-96055; Invitrogen, Thermo Fisher Scientific corporation, Massachusetts, United States; dilution, 1:1,000); and primary rabbit monoclonal antibodies against β-actin (Catalog no: M01263; Boster Biological Technology, Pleasanton, CA, United States; diluted at 1:1,000) were all identified by the antibodies used (Sigma-Aldrich, United States).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Pentagalloyl Glucose, a Major Compound in Mango Seed Kernel, Exhibits Distinct Gastroprotective Effects in Indomethacin-Induced Gastropathy in Rats via Modulating the NO/eNOS/iNOS Signaling Pathway

doi: 10.3389/fphar.2022.800986

Figure Lengend Snippet: Upper panel represents the effect of pretreatment with either pentagalloyl glucose (PGG) (100 or 200 mg/kg) or famotidine on gastric VEGF level of indomethacin-induced gastric injury. The lower panel represents densitometry analysis of the ratio of VEGF protein over β-actin protein. Data are expressed as mean ± SEM, n = 3. * and @ Statistically significant from control or indomethacin groups of rats, respectively, at p < 0.05 by one-way ANOVA was used for statistical analysis followed by Tukey’s post hoc test.

Article Snippet: Ten percent SDS-PAGE was used to separate tissue proteins, which were then transferred to nitrocellulose membranes using the Trans-Blot ® semi-dry transfer cell (Bio- Rad, Hercules, CA, United States) at 20 v for 15 min. After blocking with 1 × Tris-buffered saline/0.1% Tween 20 (TBST) with 5% non-fat dry milk for 1 h, rabbit polyclonal antibodies against VEGF (Catalog no: CAB12303; Assay genie, Dublin, Ireland; dilution, 1:500); primary rabbit polyclonal antibodies against PECAM-1 (Catalog no: PA5-96055; Invitrogen, Thermo Fisher Scientific corporation, Massachusetts, United States; dilution, 1:1,000); and primary rabbit monoclonal antibodies against β-actin (Catalog no: M01263; Boster Biological Technology, Pleasanton, CA, United States; diluted at 1:1,000) were all identified by the antibodies used (Sigma-Aldrich, United States).

Techniques: Control