primary antibodies against erk and akt  (Beyotime)

Journal: Cancer Cell International

Article Title: The anti-tumor effects of AZD4547 on ovarian cancer cells: differential responses based on c-Met and FGF19/FGFR4 expression

doi: 10.1186/s12935-024-03235-2

Figure Lengend Snippet: Inhibition of FGFR1 activation and downstream signaling by AZD4547 were dependent on the EOC cell lines and chemo-sensitive PDX. AZD4547 was applied for different durations in EOC cells. Phospho-FGFR1 and phosphorylation of downstream signaling proteins including p-AKT and p-ERK were investigated in cell lysates and tumor lysates obtained from AZD4547-treated PDX mice compared with control-treated mice by western blot analysis. β-actin was used as a loading control

Article Snippet: The blots were probed using primary antibodies against FGFR1, FGFR2 (Santa Cruz Biotechnology, USA), FGFR3, FGFR4 (Abcam), phospho-FGFR1 (p-FGFR1), total-c-Met, phospho-c-Met (p-c-Met), total-AKT, phospho-AKT (p-AKT), total-ERK, phospho-ERK (p-ERK) (Cell Signaling Technology), or β-actin (Santa Cruz Biotechnology).

Techniques: Inhibition, Activation Assay, Phospho-proteomics, Control, Western Blot

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Design, synthesis, biological evaluation and molecular docking study of 2,4-diarylimidazoles and 2,4-bis(benzyloxy)-5-arylpyrimidines as novel HSP90 N-terminal inhibitors

doi: 10.1080/14756366.2022.2124407

Figure Lengend Snippet: Western blotting assay. (a) (b) Effects of 16l (L: 6 μM; H: 18 μM) and 22k (L: 8 μM; H: 24 μM) on the expression of AKT, ERK, HSP70, and HSP90 in MCF-7 cells, with GAPDH as an internal reference. 17-AAG (15 μM) was used as a positive control. (c) (d) Statistical analysis of western blotting assays of 16l and 22k , respectively. Data are presented as the means ± SDs. * p < 0.05, ** p < 0.01, *** p < 0.001 versus control.

Article Snippet: Primary antibodies against ERK and AKT were purchased from Beyotime (Shanghai, China).

Techniques: Western Blot, Expressing, Positive Control, Control

Journal: International Journal of Oral Science

Article Title: Sphingosine-1-phosphate hinders the osteogenic differentiation of dental pulp stem cells in association with AKT signaling pathways

doi: 10.1038/s41368-022-00173-5

Figure Lengend Snippet: S1P hinders the osteogenesis of dental pulp stem cells (DPSCs) in association with a downregulation of osteogenic gene expression and AKT phosphorylation. a , b At 3 weeks after the induction of osteogenic differentiation in MC3T3-E1 cells ( a ) and DPSCs ( b ) with or without S1P (1, 5, or 10 μmol·L −1 ), each cell type was stained with alizarin red (AR) (upper). The graph demonstrates the AR-positive areas that were quantified from each individual culture plate (lower). c The transcript levels of S1PR1-5 in DPSCs at baseline were determined by qPCR analysis. d qPCR analysis of the mRNA expression of ALP, RUNX2, COL1A1, SPP1, and BGLAP2 in DPSCs treated with a vehicle or S1P (5 μmol·L −1 ) for 7 days. e DPSCs were treated with S1P (5 μmol·L −1 ) for the designated times. The protein expression of phospho-AKT (p-AKT), AKT, phospho-ERK (p-ERK), ERK, phospho-p38 (p-p38), p38, phospho-JNK (p-JNK), JNK, and β-actin was analyzed by immunoblotting (left). Densitometry quantification of p-AKT compared to AKT is represented (right); data are presented as the mean ± SEM. The experiments were at least triplicated. * P < 0.05, ** P < 0.005, *** P < 0.000 5 versus the vehicle control. P values were obtained using the Kruskal–Wallis test ( a – c ), Student’s t -test ( d ), and one-way ANOVA ( e )

Article Snippet: Non-specific interactions were blocked utilizing 5% bovine serum albumin for 1 h. The membranes were then incubated with primary antibodies against p-AKT, AKT, p-ERK, ERK, p-p38, p38, p-JNK, JNK (Cell Signaling Technology, Danvers, MA) and β-actin (Sigma-Aldrich) overnight at 4 °C, followed by incubation with the suitable secondary antibodies which were conjugated with HRP.

Techniques: Gene Expression, Phospho-proteomics, Staining, Expressing, Western Blot, Control

Journal: International Journal of Oral Science

Article Title: Sphingosine-1-phosphate hinders the osteogenic differentiation of dental pulp stem cells in association with AKT signaling pathways

doi: 10.1038/s41368-022-00173-5

Figure Lengend Snippet: S1P agonist blocks the osteogenic differentiation of DPSCs in association with a reduction in activated AKT. a DPSCs were incubated with osteogenic media to stimulate their differentiation into osteogenic cells with the designated concentrations of VPC24191 or CYM5520 for 3 weeks. AR staining was then conducted (left) and the AR-positive area was quantified from each individual culture plate (right). b DPSCs were treated with VPC24191 (10 μmol·L −1 ) or CYM5520 (10 μmol·L −1 ) in the osteogenic media for the indicated minutes. The protein expression of p-AKT, AKT, p-ERK, ERK, p-p38, p38, p-JNK, JNK, and β-actin was analyzed by immunoblotting (left). Densitometry quantification of p-AKT compared to AKT is represented (right); Data are presented as the mean ± SEM. The experiments were at least triplicated. * P < 0.05, ** P < 0.005, *** P < 0.000 5 versus vehicle control. P values were obtained using the Kruskal–Wallis test ( a ) and one-way ANOVA ( b )

Article Snippet: Non-specific interactions were blocked utilizing 5% bovine serum albumin for 1 h. The membranes were then incubated with primary antibodies against p-AKT, AKT, p-ERK, ERK, p-p38, p38, p-JNK, JNK (Cell Signaling Technology, Danvers, MA) and β-actin (Sigma-Aldrich) overnight at 4 °C, followed by incubation with the suitable secondary antibodies which were conjugated with HRP.

Techniques: Incubation, Staining, Expressing, Western Blot, Control