af3241 affinity mouse anti akt monoclonal antibody  (Bioss)

Journal: iScience

Article Title: SR9009 attenuates inflammation-related NPMSC pyroptosis and IVDD through NR1D1/NLRP3/IL-1β pathway

doi: 10.1016/j.isci.2024.109733

Figure Lengend Snippet:

Article Snippet: GSDMD , Bioss , no. bsm-33282M.

Techniques: Recombinant, RNA Sequencing Assay, Sequencing, Software, Flow Cytometry, Transmission Assay, Microscopy, CCK-8 Assay, TUNEL Assay

Journal: iScience

Article Title: SR9009 attenuates inflammation-related NPMSC pyroptosis and IVDD through NR1D1/NLRP3/IL-1β pathway

doi: 10.1016/j.isci.2024.109733

Figure Lengend Snippet:

Article Snippet: The P 3 NPMSC were fixed with paraformaldehyde (4%) for 20 min. Then they were washed twice with PBS containing 0.5% Triton X-100 for 20 min. After that the cells were incubated with 15% bovine serum albumin for 2 h at room temperature, rinsed with PBS and incubated with primary antibodies: collagen II (Bioss, China, catalog no. bs-10579R) (1:200), matrix metalloproteinase 13 (MMP-13) (Bioss, China, catalog no. bs-10240R) (1:200), NR1D1 (1:1000, Bioss, China, catalog no. bsm-33343M), NLRP3 (1:1000, Bioss, China, catalog no. bs-8878R), ASC (1:1000, Proteintech, USA, catalog no. 66444-1-Ig), caspase-1 (1:1000,abcam, China, catalog no. ab56416), IL-1β (1:1000, ABclonal, Wuhan, China, catalog no. A11025), GADPH (1:10000, ABclonal, Wuhan, China, catalog no. AC004), GSDMD (1:1000, Bioss, China, catalog no. bsm-33282M) overnight (4°C).

Techniques: Recombinant, RNA Sequencing Assay, Sequencing, Software, Flow Cytometry, Transmission Assay, Microscopy, CCK-8 Assay, TUNEL Assay

Journal: Cell Death & Disease

Article Title: Foxq1 promotes metastasis of nasopharyngeal carcinoma by inducing vasculogenic mimicry via the EGFR signaling pathway

doi: 10.1038/s41419-021-03674-z

Figure Lengend Snippet: A Immunofluorescence staining of Foxq1,EGFR,VE-cadherin,MMP2, and MMP9 in Foxq1 dysregulated 5–8F, and CNE1; Scale bars represent 50 μm. qRT-PCR (above) and western blot (down) were used to monitor the expression of EGFR signaling pathway and VM-related genes in 5–8F, and CNE1 after Foxq1 downregulation ( B , C ) or overexpression ( D , E ). F The binding motif of Foxq1 from the JASPAR database. G A diagram showing the relationship of full-length (FL) and mutant EGFR promoters. H 293 T cells were transfected with Foxq1 overexpressing or control vector. The luciferase reporter gene vectors carrying FL or EGFR promoter mutant were co-transfected respectively for 36 h. Then, the dual-luciferase activity was detected. I The CHIP-PCR assay was used to assess the binding of Foxq1 to the EGFR promoter region. J Anti-Foxq1-pulled down chromatins were analyzed by qRT-PCR. Data are presented as mean ± SD of three independent experiments.

Article Snippet: The membrane was incubated with polyclonal antibodies against EGFR (AF6043, Affinity, China), phospho-EGFR (AF3047, Affinity, China), MMP2 (AF0577, Affinity, China), MMP9 ( AF5228 , Affinity, China), VE-Cadherin (AF6265, Affinity, China), AKT (bsm-33282M, Bioss, China), phospho-AKT (bs-0876R, Bioss, China), Foxq1 (PA1-31951, Invitrogen, USA) or GAPDH (AC033, Abclonal, China) at a dilution of 1:1000, then incubated with species-specific HRP-conjugated secondary antibodies at a dilution of 1:5000.

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Binding Assay, Mutagenesis, Transfection, Plasmid Preparation, Luciferase, Activity Assay

Journal: Cell Death & Disease

Article Title: Foxq1 promotes metastasis of nasopharyngeal carcinoma by inducing vasculogenic mimicry via the EGFR signaling pathway

doi: 10.1038/s41419-021-03674-z

Figure Lengend Snippet: A VM channel formation in miR-124 overexpressing and control 5–8F cells (magnification, ×200); Scale bars represent 50 μm. B VM channel formation in 5–8F cells that overexpressed both Foxq1 and miR-124; p < 0.001. Foxq1 could significantly reverse the inhibitory effect of miR-124 on VM formation, which could be inhibited by Erlotinib (magnification, ×200); Scale bars represent 50 μm. C , D The statistical results of the average number of tubular structures in each group. E Immunofluorescence staining of Foxq1, EGFR, VE-cadherin, MMP2 and MMP9 in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively; scale bars represent 50μm. qRT-PCR ( F ) and western blot ( G ) were used to monitor the expression of EGFR signaling pathway and VM-related genes in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively. Data are presented as mean ± SD of three independent experiments.

Article Snippet: The membrane was incubated with polyclonal antibodies against EGFR (AF6043, Affinity, China), phospho-EGFR (AF3047, Affinity, China), MMP2 (AF0577, Affinity, China), MMP9 ( AF5228 , Affinity, China), VE-Cadherin (AF6265, Affinity, China), AKT (bsm-33282M, Bioss, China), phospho-AKT (bs-0876R, Bioss, China), Foxq1 (PA1-31951, Invitrogen, USA) or GAPDH (AC033, Abclonal, China) at a dilution of 1:1000, then incubated with species-specific HRP-conjugated secondary antibodies at a dilution of 1:5000.

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot, Expressing