Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation 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: Biomedicines

Article Title: Blockade of the SRC/STAT3/BCL-2 Signaling Axis Sustains the Cytotoxicity in Human Colorectal Cancer Cell Lines Induced by Dehydroxyhispolon Methyl Ether

doi: 10.3390/biomedicines11092530

Figure Lengend Snippet: Inhibition of SRC activation underlies DHME-induced STAT3 blockade. ( A ) Limited effect of DHME on JAK2 activation. Human CRC cells treated with DHME (0, 20, 40 μM) were subjected to immunoblotting 24 h later for the amounts of tyrosine 1007/1008-dual phosphorylated JAK2 (p-JAK2 (Y1007/Y1008)), a surrogate marker of JAK2 activation. ( B ) DHME suppresses the activation of SRC. DHME-treated human CRC cells were subject to immunoblotting for the levels of tyrosine 406-phosphorylated SRC (p-SRC (Y406)), a substitute marker of SRC activation. ( C ) Sustained SRC activation thwarts DHME-induced blockade of STAT3 activation and apoptosis. Human CRC stable clones of v-src (a dominant-active SRC) or its vector control were subject to 24 h treatment with DHME and then were underwent immunoblotting for the levels of p-STAT3 (Y705) and cleaved PARP (c-PARP).

Article Snippet: Primary antibodies against cleaved PARP (#9541), HA-tag (#3724), phospho-STAT3 (Y705) (#9145), phospho-JAK2 (Y1007/1008) (#3776), JAK2 (#3230), phospho-Src (Y416) (#6743), and Src (#2108) were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Inhibition, Activation Assay, Western Blot, Marker, Clone Assay, Plasmid Preparation, Control

Journal: American Journal of Translational Research

Article Title: Novel apoE receptor mimetics reduce LPS-induced microglial inflammation

doi:

Figure Lengend Snippet: 6KApoEp attenuates LPS-induced JAK2 and STAT3 phosphorylation. BV2 cells were treated with LPS at 100 ng/ml in the absence or presence of 6KApoEp at 10 µM for 0, 15, 30 or 60 min. Total (JAK2 and STAT3) and phosphorylated JAK2 and STAT3 (p-JAK2 and p-STAT3) in cell lysates was then determined by WB, utilizing anti-total JAK2 (Cell Signaling, #3230), anti-total STAT3 (#9132), anti-phospho-JAK2 (#3774) and anti-phospho STAT3 antibodies (#9131). Representative blots are shown for each condition and band density ratios of p-JAK2 to JAK2 and p-STAT3 to STAT3 determined by densitometry analysis are shown below each blot. Full non-adjusted images of WB shown in Figure S2. LPS-induced JAK2 and STAT3 phosphorylation was reduced by 6KApoEp between 15 and 60 min of treatment. Total JAK2 and STAT3 levels were similar across treatment conditions. Results are expressed as mean ± S.E.M. Asterisk indicate P < 0.05 compared with LPS alone.

Article Snippet: BV2 cells were treated with LPS at 100 ng/ml in the absence or presence of 6KApoEp at 10 µM for 0, 15, 30 or 60 min. Total (JAK2 and STAT3) and phosphorylated JAK2 and STAT3 (p-JAK2 and p-STAT3) in cell lysates was then determined by WB, utilizing anti-total JAK2 (Cell Signaling, #3230), anti-total STAT3 (#9132), anti-phospho-JAK2 (#3774) and anti-phospho STAT3 antibodies (#9131).

Techniques:

Journal:

Article Title: Prolactin and ErbB4/HER4 Signaling Interact via Janus Kinase 2 to Induce Mammary Epithelial Cell Gene Expression Differentiation

doi: 10.1210/me.2008-0055

Figure Lengend Snippet: ErbB4 Is Required for JAK2 Activation in Response to HB-EGF But Not PRL

Article Snippet: Western Analysis and Immunoprecipitation Cells were lysed, and proteins were precipitated with the following antibodies as described previously ( 57 ): rabbit polyclonal anti-ErbB1 and -ErbB4 (C terminus) generated in this laboratory and previously described ( 57 ); JAK2, PRLR, phosphotyrosine (PY20), Src, and P-Src (Santa Cruz Biotechnologies, Santa Cruz, CA), STAT5A and P-Tyr 694 STAT5 (Zymed Laboratories, San Francisco, CA), p44/42, and phospho-p44/42 (Cell Signaling Technologies, Beverly, MA).

Techniques: Activation Assay

Journal:

Article Title: Prolactin and ErbB4/HER4 Signaling Interact via Janus Kinase 2 to Induce Mammary Epithelial Cell Gene Expression Differentiation

doi: 10.1210/me.2008-0055

Figure Lengend Snippet: Kinase Activity and Expression of JAK2 Is Required for ErbB4-Mediated STAT5A Activation

Article Snippet: Western Analysis and Immunoprecipitation Cells were lysed, and proteins were precipitated with the following antibodies as described previously ( 57 ): rabbit polyclonal anti-ErbB1 and -ErbB4 (C terminus) generated in this laboratory and previously described ( 57 ); JAK2, PRLR, phosphotyrosine (PY20), Src, and P-Src (Santa Cruz Biotechnologies, Santa Cruz, CA), STAT5A and P-Tyr 694 STAT5 (Zymed Laboratories, San Francisco, CA), p44/42, and phospho-p44/42 (Cell Signaling Technologies, Beverly, MA).

Techniques: Activity Assay, Expressing, Activation Assay