phosphorylated jak2 p jak2  (Cell Signaling Technology Inc)

Journal: Journal of Thoracic Disease

Article Title: Mebendazole induces apoptosis and inhibits migration via the reactive oxygen species-mediated STAT3 signaling downregulation in non-small cell lung cancer

doi: 10.21037/jtd-23-1978

Figure Lengend Snippet: MBZ inhibited the ROS-JAK2-STAT3 signaling cascade in A549 and H460 cells. (A) The protein levels of JAK2-STAT3 signaling cascade treated with MBZ (0.5 µM) were investigated with Western blotting in A549 and H460 cells. (B) STAT3 Luci and β-gal plasmids were cotransfected into A549 and H460 cells. The cells were then administered MBZ alone or in combination with NAC for 48 h, which was followed by luciferase assays. Data from three independent assays were pooled. *, P<0.05; **, P<0.01; ***, P<0.001. MBZ, mebendazole; ROS, reactive oxygen species; JAK2, Janus kinase 2; STAT3, signal transducer and activator of the transcription 3; NAC, N-acetylcysteine.

Article Snippet: Stattic was purchased from Selleck. β-actin (#3700), β-tubulin (#2128), E-Cadherin (#14472), N-Cadherin (#13116), ADP-ribose polymerase (PARP; #9532), JAK2 (#3230), caspase 3 (#9662), STAT3 (#9139), cleaved caspase 3 (#9661), C-MYC (#18583), phosphorylated JAK2 (p-JAK2) (#66245), Bcl-xl (#2764), SNAI1 (#3879), TWIST1 (#69366), and phosphorylated STAT3 (Tyr705) (p-STAT3) (#9145) antibodies were acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Luciferase

Journal: Brain sciences

Article Title: Targeting Microglia/Macrophages Notch1 Protects Neurons from Pyroptosis in Ischemic Stroke.

doi: 10.3390/brainsci13121657

Figure Lengend Snippet: Figure 6. Notch1 knockdown in microglial BV2 cells inhibits OGD/R-induced JAK2/STAT3 activation and pyroptosis in HT22 cells. (A) Western blot analysis of the expression of pyroptosis-associated proteins in HT22 cells co-cultured with NC-siRNA-transfected BV2 cells and HT22 cells co-cultured with Notch1-siRNA-transfected BV2 cells. (B) Relative quantification of pyroptosis-associated protein expression in HT22 cells. (C) mRNA expression levels of inflammasome-associated proteins (NLRP3 and Cl-caspase1) and pyroptosis-associated proteins (Cl-GSDMD, IL-1β and IL-18) in HT22 cells co- cultured with NC-siRNA-transfected BV2 cells and those co-cultured with Notch1-siRNA-transfected BV2 cells after OGD/R or sham treatment. (D) Protein–protein interaction (PPI) network of Notch1 and JAK2/STAT3. (E) Representative Western blot images of P-JAK2, JAK2, P-STAT3, and STAT3 in HT22 cells co-cultured with NC-siRNA-transfected BV2 cells and HT22 cells co-cultured with Notch1- siRNA-transfected BV2 cells after OGD/R or sham treatment. (F) Relative quantification of P-JAK2 and P-STAT3 protein expression. Mean ± SD. n = 5 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were then incubated overnight at 4 ◦C with primary antibodies specific for Notch1 (diluted 1:1000; MA5-32080, Invitrogen), cleaved caspase1 (diluted 1:1000; 89332, Cell Signaling Technology, Danvers, MA, USA), cleaved GSDMD (diluted 1:1000; P30823S, ABMART, Shanghai, China), NLRP3 (diluted 1:1000; T55651S, ABMART, Shanghai, China), IL1β (diluted 1:1000; 12242, Cell Signaling Technology), IL18 (diluted 1:1000; M027287S, ABMART), JAK2 (diluted 1:1000; 3230, Cell Signaling Technology, Danvers, MA, USA), P-JAK2 (diluted 1:1000; 66245, Cell Signaling Technology), STAT3 (diluted 1:1000; 12640S, Cell Signaling Technology Danvers, MA, USA), P-STAT3 (diluted 1:1000; 9145S, Cell Signaling Technology, Danvers, MA, USA), Bax (diluted 1:1000; 41162, Cell Signaling Technology, Danvers, MA, USA), Bcl2 (diluted 1:1000; sc-492, Santa Cruz Biotechnology, Dallas, TX, USA), iNOS (diluted 1:1000; ab49999, Abcam, Cambridge, MA, USA), GAPDH (diluted 1:1000; GB11002, Servicebio, Wuhan, China) and β-actin (diluted 1:1000; GB12001, Servicebio, Wuhan, China).

Techniques: Knockdown, Activation Assay, Western Blot, Expressing, Cell Culture, Transfection, Quantitative Proteomics