other:Article Title: Selective inhibition of long isoforms of phosphodiesterase 4D mitigates liver fibrosis in mouse models
Article Snippet: Primary antibodies were used at 1:1,000 dilution, including COL1A1 (#84336, Cell signaling); ACTA2 (#A2547, Sigma-Aldrich); COL1A1 (#84336, Cell signaling); p-SMAD2 (#3108, Cell signaling); p-SMAD3 (#ab52903, abcam); SMAD2/3 (#8685, Cell signaling); SMAD4 (#46535, Cell signaling); LAMIN (#13435, Cell signaling); p-ERK (#4370, Cell signaling); ERK (#4696, Cell signaling); p-IKK (#2697, Cell signaling); IKK (#11930, Cell signaling); p-NF-κB p65 S536 (#3033, Cell signaling); NF-κB P65 (#8242, Cell signaling); p-AKT (#4060, Cell signaling); AKT (#4691, Cell signaling); p-FOXO1 S256 (#9461, Cell signaling); p-FOXO3a S253 (#13129, Cell signaling); FOXO1 (#2880, Cell signaling); p18 INK4C (#2896, Cell signaling); p21 Waf1/Cip1 (#2947, Cell signaling); CDK4 (#12790, Cell signaling); CDK6 (#3136, Cell signaling); PARP (#9542, Cell signaling); CASPASE3 (#14220, Cell signaling); Integrin a5 (#ab150361, abcam); p-FAK Y397 (#8556, Cell signaling); p-FAK Y925 (#3284, Cell signaling); FAK (#13009, Cell signaling); FLAG (#F1804, Sigma-Aldrich); a-tubulin (sc-8035, Santa Cruz).
Article Title: Selective inhibition of long isoforms of phosphodiesterase 4D mitigates liver fibrosis in mouse models
Article Snippet: COL1A1 (#84336, Cell signaling); ACTA2 (#A2547, Sigma-Aldrich); COL1A1 (#84336, Cell signaling); p-SMAD2 (#3108, Cell signaling); p-SMAD3 (#ab52903, abcam); SMAD2/3 (#8685, Cell signaling); SMAD4 (#46535, Cell signaling); LAMIN (#13435, Cell signaling); p-ERK (#4370, Cell signaling); ERK (#4696, Cell signaling); p-IKK (#2697, Cell signaling); IKK (#11930, Cell signaling); p-NF-κB p65 S536 (#3033, Cell signaling); NF-κB P65 (#8242, Cell signaling); p-AKT (#4060, Cell signaling); AKT (#4691, Cell signaling); p-FOXO1 S256 (#9461, Cell signaling); p-FOXO3a S253 (#13129, Cell signaling); FOXO1 (#2880, Cell signaling); p18 INK4C (#2896, Cell signaling); p21 Waf1/Cip1 (#2947, Cell signaling); CDK4 (#12790, Cell signaling); CDK6 (#3136, Cell signaling); PARP (#9542, Cell signaling); CASPASE3 (#14220, Cell signaling); Integrin a5 (#ab150361, abcam); p-FAK Y397 (#8556, Cell signaling); p-FAK Y925 (#3284, Cell signaling); FAK (#13009, Cell signaling); FLAG (#F1804, Sigma-Aldrich); a-tubulin (sc-8035, Santa Cruz).
Article Title: Apelin promotes RANKL‑mediated osteoclastogenesis by activating MAPK and NF‑κB pathways.
Article Snippet: The JnK inhibitor SP600125 was purchased from enzo life Sciences, inc. whereas the p38 inhibitor SB203580 and nF‐κB inhibitor pyrrolidinedithiocarbamate ammonium (PdTc) were obtained from MilliporeSigma. antibodies against iKK (cat. no. GTX52348), iκB (cat. no. GTX110521), p65 (cat. no. GTX102090) and β‐actin (1:10,000; cat. no. GT5512) were purchased from GeneTex international corporation. antibodies for p‐iKK (cat. no. 2697S) were from cell Signaling Technology, inc., antibodies for p‐iκB (cat. no. sc‐8404) were obtained from Santa cruz Biotechnology, inc. and antibodies targeting p‐p65 (cat. no. aP0124) were obtained from aBclonal Biotech co., ltd.
Western Blot:Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis
Article Snippet: .. Western blot antibodies include monoclonal mouse antiβ-actin (1:5000 dilution, Cat# AF7018, Affinity), PI3K (Cat# 4257, CST), P-PI3K (Cat# 4228, CST), AKT (Cat# 4685, CST), P-AKT (Cat# 4060, CST), IKKa (Cat# 11930, CST), IKKb (Cat# 8943, CST), p-IKK (Cat# 2697, CST), pP65 (Cat# 3033, CST), P65 (Cat# 8242, CST), IκBa (Cat# 4814, CST), p-IκBa (Cat# 2859, CST), GAPDH (Cat# 5174, CST), PDPN (Cat# sc-376695, Santa Cruz) at 1:1000 dilution, and secondary antibody-goat anti-Rabbit and Mouse (Cat# M21003, Abmart). .. Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), humanCCL2 (Cat# 300-04-5, PeproTech).
Article Title: PDPN+ cancer‐associated fibroblasts enhance gastric cancer angiogenesis via AKT/NF‐κB activation and the CCL2‐ACKR1 axis
Article Snippet: .. Western blot antibodies include monoclonal mouse anti‐β‐actin (1:5000 dilution, Cat# AF7018, Affinity), PI3K (Cat# 4257, CST), P‐PI3K (Cat# 4228, CST), AKT (Cat# 4685, CST), P‐AKT (Cat# 4060, CST), IKKa (Cat# 11930, CST), IKKb (Cat# 8943, CST), p‐IKK (Cat# 2697, CST), p‐P65 (Cat# 3033, CST), P65 (Cat# 8242, CST), IκBa (Cat# 4814, CST), p‐IκBa (Cat# 2859, CST), GAPDH (Cat# 5174, CST), PDPN (Cat# sc‐376695, Santa Cruz) at 1:1000 dilution, and secondary antibody‐goat anti‐Rabbit and Mouse (Cat# M21003, Abmart). .. Other antibodies include CD31 (Cat# ab81289, Abcam), CCL2 neutralizing antibody (Cat# MAB679, R&D), human CCL2 (Cat# 300‐04‐5, PeproTech).
Article Title: Hypoxia-preconditioned cardiomyocyte-derived extracellular vesicles alleviate myocardial ischemic injury by reprogramming macrophage polarization via the Fgl2/NF-κB pathway.
Article Snippet: Protein detection was performed using an enhanced chemiluminescence (ECL) system, and results were analyzed using image system software (Protein Simple, USA). .. Primary antibodies used for Western blot included Fgl2 (Proteintech, 11827–1-AP), TLR4 (Proteintech, 66350–1-Ig), iNOS (Proteintech, 22226–1-AP), COX2 (Proteintech, 12375–1-AP), p-IKK (CST #2697), IKK (CST #8943), p-IκB (Proteintech, 82349–1-RR), IκB (Proteintech, 10268–1-AP), p-NF-κB (Proteintech, 82335–1-RR), NF-κB (Proteintech, 10745–1-AP), p-MAPK (Proteintech, 28796–1-AP), MAPK (Proteintech, 14064–1-AP), Bax (Proteintech, 50599–2-Ig), Bcl2 (Proteintech, 26593–1-AP), TNF-α (Proteintech, 17590–1-AP), IL-1β (Abcam, 254360), IL-6 (Proteintech, 21865–1-AP), IL-10 (Proteintech, 60269–1- Ig), Arg-1 (Proteintech, 16001–1-AP), APOA1 (Proteintech, 14427–1- AP), β-actin (HUABIO, EM21002), and GAPDH (Proteintech, 60004–1- Ig). .. The phagocytic capacity of RAW264.7 macrophages was assessed using the VybrantTM Phagocytosis Assay Kit (V-6694, Molecular Probes, USA).
Incubation:Article Title: CircFRRS1 drives neuroinflammation through the miR-27a-3p/TLR4 pathway after deep hypothermic circulatory arrest
Article Snippet: .. After the primary antibody protocol was completed, mMembranes were then washed in TBST and incubated with secondary antibody at room temperature for 1 h. The primary and secondary antibodies used in this experiment were as follows: primary antibodies: TLR4 (1:1,000; GB11519, Servicebio), Myd88 (1:1,000; GB111554 , Servicebio), p-IKK (1:1,000; CST#2697, Cell Signaling Technology), IKK (1:1,000; WL01900, Wanleibio), p-IκB (1:1,000; CST #2859, Cell Signaling Technology), IκB (1:1,000; CST#4812, Cell Signaling Technology), p-P65(1:500; CST#3033, Cell Signaling Technology), P65 (1:1,000; CST#3034, Cell Signaling Technology) and NLRP3 (1:1,000; GB114320 , Servicebio); secondary antibodies: HRP conjugated Goat Anti-Rabbit IgG (H+L) (GB23303, Servicebio). .. Bound proteins were detected using Amersham Imager 800 (Cytiva, Inc.) and quantified by ImageJ software (v1.54). β-Actin was used as an internal control for protein inputs.
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