Journal: The Journal of Cell Biology

Article Title: α-Syntrophin regulates ARMS localization at the neuromuscular junction and enhances EphA4 signaling in an ARMS-dependent manner

doi: 10.1083/jcb.200412008

Figure Lengend Snippet: ARMS and α-syntrophin enhanced the EphA4-induced Jak/Stat activation. (A) COS7 cells were transfected with EphA4, EphA4 KD mutant, or ARMS, and cell lysate was immunoblotted with antibodies against phosphorylated Tyr 1007/1008 of mouse Jak2, Tyr 1054/1055 of human Tyk2, Tyr 701 of Stat1, and Thr 202 /Tyr 204 of ERK1/2. p-Tyr; phosphorylated Tyr. (B) Quantification of the Western blots. y axis represents the fold change of tyrosine phosphorylation. n = 3; *, P < 0.05; **, P < 0.005. (C) COS7 cells were transfected with EphA4, ARMS, α-syntrophin, or α-syntrophin mPDZ, and cell lysate was immunoblotted with antibodies against phosphorylated Tyr 1007/1008 of mouse Jak2, Tyr 1054/1055 of human Tyk2, Tyr 701 of Stat1, and Thr 202 /Tyr 204 of ERK1/2. (D) Quantification of the Western blots. n = 3; *, P < 0.05; **, P < 0.005. Error bars represent the SEM.

Article Snippet: Polyclonal anti-HA antibodies, EphA4 antibody (sc-921), and Jak2 antibody (sc-278) were purchased from Santa Cruz Biotechnology, Inc., and the mAb that recognizes the α-isoform of Stat1 was obtained from Zymed Laboratories.

Techniques: Activation Assay, Transfection, Mutagenesis, Western Blot, Phospho-proteomics

Journal: Neoplasia (New York, N.Y.)

Article Title: INCB16562, a JAK1/2 Selective Inhibitor, Is Efficacious against Multiple Myeloma Cells and Reverses the Protective Effects of Cytokine and Stromal Cell Support

doi:

Figure Lengend Snippet: Selective Inhibition of JAKs by INCB16562.

Article Snippet: The primary antibodies specific for the following proteins were used at the indicated dilutions: phospho-STAT3 (1:1000), STAT3 (1:1000), STAT5 (1:1000), phospho-JAK2 (1:1000), and JAK2 (1:1000; all from Cell Signaling, Beverly, MA); phospho-STAT5 (1:1000; Millipore, Temecula, CA); Mcl-1 (1:100), poly (ADP-ribose) polymerase (PARP; 1:100), Bcl-2 (1:100), Bcl-X L (1:100), α-actin (1:100; all from Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Inhibition

Journal: Neoplasia (New York, N.Y.)

Article Title: INCB16562, a JAK1/2 Selective Inhibitor, Is Efficacious against Multiple Myeloma Cells and Reverses the Protective Effects of Cytokine and Stromal Cell Support

doi:

Figure Lengend Snippet: INCB16562 Is a Potent JAK1/2 Inhibitor.

Article Snippet: The primary antibodies specific for the following proteins were used at the indicated dilutions: phospho-STAT3 (1:1000), STAT3 (1:1000), STAT5 (1:1000), phospho-JAK2 (1:1000), and JAK2 (1:1000; all from Cell Signaling, Beverly, MA); phospho-STAT5 (1:1000; Millipore, Temecula, CA); Mcl-1 (1:100), poly (ADP-ribose) polymerase (PARP; 1:100), Bcl-2 (1:100), Bcl-X L (1:100), α-actin (1:100; all from Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques:

Journal: Molecular Neurobiology

Article Title: Melatonin Reduces Neuroinflammation and Improves Axonal Hypomyelination by Modulating M1/M2 Microglia Polarization via JAK2-STAT3-Telomerase Pathway in Postnatal Rats Exposed to Lipopolysaccharide

doi: 10.1007/s12035-021-02568-7

Figure Lengend Snippet: Primary antibodies used in experiments

Article Snippet: P-JAK2 , Rabbit , Cell Signaling Technology , 3776 , WB (1:1000).

Techniques: Concentration Assay

Journal: Molecular Neurobiology

Article Title: Melatonin Reduces Neuroinflammation and Improves Axonal Hypomyelination by Modulating M1/M2 Microglia Polarization via JAK2-STAT3-Telomerase Pathway in Postnatal Rats Exposed to Lipopolysaccharide

doi: 10.1007/s12035-021-02568-7

Figure Lengend Snippet: Melatonin activates JAK2/STAT3 pathway in microglia exposed to LPS in vitro. Panel A shows p-JAK2(131 kDa), JAK2(131 kDa), p-STAT3(88 kDa), STAT3(88 kDa) and β-actin(42 kDa) immunoreactive bands. Bar graphs in B - E show optical density changes of p-JAK2, JAK2, p-STAT3, STAT3 relative to β-actin of each group. Note melatonin treatment activates JAK2/STAT3 pathway and then modulate the conversion of M1 to M2 polarization. Note the effect of melatonin was blocked by luzindole. Scale bars: A – I 20 µm. * P < 0.05, n = 5 for each group

Article Snippet: P-JAK2 , Rabbit , Cell Signaling Technology , 3776 , WB (1:1000).

Techniques: In Vitro

Journal: Molecular Neurobiology

Article Title: Melatonin Reduces Neuroinflammation and Improves Axonal Hypomyelination by Modulating M1/M2 Microglia Polarization via JAK2-STAT3-Telomerase Pathway in Postnatal Rats Exposed to Lipopolysaccharide

doi: 10.1007/s12035-021-02568-7

Figure Lengend Snippet: Table of Contents Image (TOCI): a schematic diagram depicting the cellular and molecular events associated with melatonin treatment in postnatal rats given LPS injection. The illustration follows two paths: the solid line shows that melatonin binds to its cognate receptor (MT1) on the microglia which activates the JAK2/STAT3 pathways and increases the expression of telomerase in the nucleus. This decreases M1 microglial production of proinflammatory cytokines, such as IL-1β, TNF-α and iNOS, and increases M2 production of anti-inflammatory cytokines, such as CD206 and TGF-β. Dash line denotes the effect of LPS on microglia. LPS stimulates toll-like receptor 4 (TLR4) which recruits its downstream NF-κB pathway, and ultimately mediates the production of proinflammatory mediators . However, LPS inhibits the activation of JKA2/STAT3 pathway and reduces the expression p-JAK2 and p-STAT3 proteins expression, which is contrary to melatonin treatment. Production of proinflammatory cytokines by M1 microglia causes hypomyelination in the corpus callosum after LPS administration; while production of anti-inflammatory cytokines by M2 induced by melatonin improves hypomyelination

Article Snippet: P-JAK2 , Rabbit , Cell Signaling Technology , 3776 , WB (1:1000).

Techniques: Injection, Expressing, Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Tristetraprolin-dependent Post-transcriptional Regulation of Inflammatory Cytokine mRNA Expression by Apolipoprotein A-I

doi: 10.1074/jbc.M110.202275

Figure Lengend Snippet: STAT3 activation is involved in the apoA-I-mediated increase of TTP expression in LPS-treated macrophages. A, THP-1 macrophages were treated with apoA-I for different times as indicated. Proteins were extracted, and the phosphorylated STAT1, STAT3, and STAT6 levels were measured by immunoblot analyses with antibodies specific for phosphorylated signal transducer and activator of transcriptions. Values were normalized against β-actin. *, p < 0.05 versus 0 min. B, THP-1 macrophages were treated with apoA-I for different times as indicated. The nuclear proteins extracted from cells were subjected to immunoblot analyses with antibodies against STAT3 and histone H1. *, p < 0.05 versus 0 min. C, THP-1 macrophages were pretreated with apoA-I for 30 min. Thereafter, the medium was replaced with fresh medium containing the AG-490 (30 μm). Then cells were incubated for another 30 min, and total or nuclear proteins were subjected to immunoblot analyses with antibody against p-JAK2 and STAT3. *, p < 0.05. D, THP-1 macrophages were transfected with control (WT) or STAT3 siRNA for 48 h, and protein samples were immunoblotted with STAT3 and β-actin antibodies. *, p < 0.05 versus control group. E, THP-1 macrophages transfected with control or STAT3 siRNA were incubated with LPS for 4 h with or without pretreatment of apoA-I. Protein samples were immunoblotted with TTP and β-actin antibodies. *, p < 0.05 versus control group. F, THP-1 macrophages were pretreated with AG-490 or DMSO for 1 h, and cells were then incubated with LPS for another 4 h with or without pretreatment of apoA-I. Protein samples were immunoblotted with TTP and β-actin antibodies. *, p < 0.05 compared with DMSO group. All of the results are the mean ± S.D. of quadruplicate values from three separate experiments.

Article Snippet: Antibodies for phosphorylation and total STAT3 and phosphor- JAK2 were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Expressing, Western Blot, Incubation, Transfection, Control

Journal: The Journal of Biological Chemistry

Article Title: Tristetraprolin-dependent Post-transcriptional Regulation of Inflammatory Cytokine mRNA Expression by Apolipoprotein A-I

doi: 10.1074/jbc.M110.202275

Figure Lengend Snippet: Schematic representation of the effects of apoA-I on TNF-α mRNA stabilization in LPS-stimulated macrophages. The results of the present studies revealed the following scheme for the possible mechanisms: apoA-I down-regulates the expression of TNF-α via a post-transcriptional regulation manner in macrophages; when macrophages are treated with apoA-I, the JAK2/STAT3 signaling pathway is activated, and the STAT3 dimers bind to the regulated elements of TTP in the nucleus. Then the expression of TTP is up-regulated, which in turn promotes the degradation of inflammatory cytokine mRNA through its 3′-UTR AREs. Plus sign indicates activation; scissors indicate degradation.

Article Snippet: Antibodies for phosphorylation and total STAT3 and phosphor- JAK2 were purchased from Cell Signaling Technology.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: LncTUG1 promotes hepatocellular carcinoma immune evasion via upregulating PD-L1 expression

doi: 10.1038/s41598-023-42948-8

Figure Lengend Snippet: TUG1 affects the JAK2/ STAT3 pathway to regulate PD-L1. ( A ): The relative mRNA expression of JAK2 was analyzed in shTUG1 HCC cells. ( B ): The relative mRNA expression of STAT3 was analyzed in shTUG1 HCC cells. ( C ): The protein levels of pJAK2, JAK2, pSTAT3, STAT3, PD-L1 were analyzed in shTUG1 HCC cells by western blotting. ( D ) pJAK2, JAK2,and PDL1 protein levels are assessed by immunoblotting in HCC-LM3 cell was transfected with TUG1 silenced (shTUG1) and/or JAK2 overexpressed (JAK2-OE). ( E ) pSTAT3,STAT3,and PDL1 protein levels are assessed by immunoblotting in HCC-LM3 cell was transfected with TUG1 silenced (shTUG1) and/or STAT3 overexpressed (STAT3-OE).

Article Snippet: Protein detection was performed by using the following primary antibodies: anti-PD-L1 (ab205921, Abcam), anti-GAPDH (ab8245, Abcam), anti-JAK2 (#3230, Cell Signaling Technology), anti-pJAK2 Tyr1007/1008 (#3771,Cell Signaling Technology),anti-STAT3 (#9139, Cell Signaling Technology), anti-pSTAT3 Tyr705 (#9145, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Transfection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sterol Regulatory Element-Binding Protein-1c Regulates Inflammasome Activation in Gingival Fibroblasts Infected with High-Glucose-Treated Porphyromonas gingivalis

doi: 10.3389/fcimb.2016.00195

Figure Lengend Snippet: JAK2 is required for Akt activation and SREBP-1c-mediated NLRP3 and IL-1β expression . HGFs were kept as CL or infected by HGPg (20 MOI). Before being kept as CL or infected by HGPg, HGFs were transfected with si-JAK1, 2, or 3. (A) The phosphorylation of Akt and p70S6K in HGF cell lysate after HGPg infection was determined using Western blotting. (B) The expression of NLRP3 and SREBP-1c in HGF cell lysate after HGPg infection was determined using Western blotting. (C) IL-1β secretion in medium was determined by ELISA analyses. The results are shown as mean ± SEM. * P < 0.05 vs. CL. # P < 0.05 vs. si-CL-transfected HGFs with HGPg infection. (D) The phosphorylation of JAK2 in HGF cell lysate after HGPg infection was determined using Western blotting.

Article Snippet: Rabbit polyclonal antibodies against phospho-p70S6K, p70S6K, phospho-JAK2, and JAK2 were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Activation Assay, Expressing, Infection, Transfection, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sterol Regulatory Element-Binding Protein-1c Regulates Inflammasome Activation in Gingival Fibroblasts Infected with High-Glucose-Treated Porphyromonas gingivalis

doi: 10.3389/fcimb.2016.00195

Figure Lengend Snippet: β1 integrin is required for HGPg-induced NLRP3 expression . HGFs were kept as CL or infected by HGPg (20 MOI). Before being kept as CL or infected by HGPg, HGFs were transfected with si-integrin β1 or β2, or pretreated with neutralizing antibody against integrin β1. (A) The phosphorylation of JAK2 in HGF cell lysate after HGPg infection was determined using Western blotting. (B) The phosphorylation of Akt and p70S6K, and the expression of NLRP3 and SREBP-1c in HGF cell lysate after HGPg infection, were determined using Western blotting. (C) IL-1β secretion in medium was determined by ELISA analyses. The results are shown as mean ± SEM. * P < 0.05 vs. CL. # P < 0.05 vs. si-CL-transfected or IgG-treated HGFs with HGPg infection.

Article Snippet: Rabbit polyclonal antibodies against phospho-p70S6K, p70S6K, phospho-JAK2, and JAK2 were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Infection, Transfection, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay