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: Communications biology

Article Title: S100A8/A9 promotes endometrial fibrosis via regulating RAGE/JAK2/STAT3 signaling pathway.

doi: 10.1038/s42003-024-05814-5

Figure Lengend Snippet: Fig. 4 S100A8/A9 activates the JAk2/STAT3 signaling pathway through RAGE. a–c The expression of JAK2, p-JAK2, STAT3, and p-STAT3 was detected by western blot after treating hEnSCs with S100A8/A9 for 0.5 h, 1 h, 3 h, and 6 h, n = 6 per group, ordinary one-way ANOVA with Dunnett’s multiple comparison test. d–f Pretreating hEnSCs with AG490 for 1 h followed by S100A8/A9 treatment for 24 h and 48 h, the expression of Col1 and α- SMA was detected by western blot, n = 6 per group, ordinary one-way ANOVA with Tukey’s multiple comparison test. g–i Pretreating hEnSCs with FPS- ZM1 or TAK-242 for 1 h followed by S100A8/A9 stimulation for 6 h, the expression of JAK2, p-JAK2, STAT3, and p-STAT3 was detected by western blot, n = 6 per group, ordinary one-way ANOVA with Dunnett’s multiple comparison test. j Immunofluorescence was used to detect the expression of STAT3 in hEnSCs. red: STAT3, blue: DAPI, scale bar = 50 μm. k Representative immunohistochemistry images of RAGE, p-JAK2, p-STAT3 in endometrial tissue sections from normal subjects and patients with IUA are shown, scale bar = 100 μm. Data are presented as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 compared with indicated groups. ns, not significant (P > 0.05).

Article Snippet: A quantity of 10 μg of protein was loaded onto a 10% gradient polyacrylamide gel, electrophoretically transferred to a polyvinylidene difluoride membrane and probed with the following primary antibodies: S100A8/A9 (1:1000, Abcam, UK), Col1 (1:2000, Proteintech, China), α-SMA (1:6000, Proteintech, China), JAK2 (1:1000, Proteintech, China), p-JAK2 (Tyr1007) (1:1000, Absin, China), STAT3 (1:2000, Proteintech, China), pSTAT3(Tyr705) (1:1000, Absin, China), RAGE(1:1000, Proteintech, China), TLR4 (1:1000, Proteintech, China), and GAPDH (1:1000, Absin, China), which was used as an internal control.

Techniques: Expressing, Western Blot, Comparison, Immunohistochemistry

Journal: Communications biology

Article Title: S100A8/A9 promotes endometrial fibrosis via regulating RAGE/JAK2/STAT3 signaling pathway.

doi: 10.1038/s42003-024-05814-5

Figure Lengend Snippet: Fig. 9 Schematic representation of the role of S100A8/A9 in activating hEnSCs. Following endometrial injury, neutrophils migrate to the trauma site, releasing S100A8/A9. S100A8/A9 binds to the extracellular domains of RAGE on the membrane of hEnSCs, activating the JAK2/STAT3 signaling pathway. Subsequently, it promotes the differentiation of hEnSCs into myofibroblasts, with increased expression of Col1 and α-SMA proteins.

Article Snippet: A quantity of 10 μg of protein was loaded onto a 10% gradient polyacrylamide gel, electrophoretically transferred to a polyvinylidene difluoride membrane and probed with the following primary antibodies: S100A8/A9 (1:1000, Abcam, UK), Col1 (1:2000, Proteintech, China), α-SMA (1:6000, Proteintech, China), JAK2 (1:1000, Proteintech, China), p-JAK2 (Tyr1007) (1:1000, Absin, China), STAT3 (1:2000, Proteintech, China), pSTAT3(Tyr705) (1:1000, Absin, China), RAGE(1:1000, Proteintech, China), TLR4 (1:1000, Proteintech, China), and GAPDH (1:1000, Absin, China), which was used as an internal control.

Techniques: Membrane, Expressing

Journal: Poultry Science

Article Title: Characterization and functional analyses of novel chicken leukocyte immunoglobulin-like receptor subfamily B members 4 and 5

doi: 10.3382/ps/pez442

Figure Lengend Snippet: LILRB4R, -B4S, -B5R, and -B5S glycoprotein regulated the JAK-STAT signaling pathway. ( A ) Western blotting results of JAK2/TYK2, STAT1/3, and SOCS1 after LILRB4–5 transfection of HD11 cell line. ( B ) Changes in mRNA levels of JAK2/TYK2, STAT1/3 and SOCS1 genes after LILRB4–5 transfection in HD11 cell line were detected by qRT-PCR. Data are presented as the mean ± SEM ( n = 3) of 3 independent experiments: * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The following reagents were from the indicated manufacturers: mouse anti-chicken MHC Class I-PE and mouse anti-chicken β2m-PE antibody (Southern Biotech, Birmingham, AL, USA); rabbit anti-chicken STAT1 (phospho-Ser 727 ), anti-chicken STAT3 (phospho-Ser 727 ), and anti-chicken JAK2 (phospho-Tyr 1007 /Tyr 1008 ) antibody (Santa Cruz Biotech Dallas, Texas, USA); rabbit anti-chicken SOCS1, anti-chicken STAT1, anti-chicken STAT3 antibodies, horseradish peroxidase ( HRP )-linked anti-rabbit secondary antibodies, and protein G–sepharose bead (Sigma-Aldrich, Louis, MO, USA); rabbit anti-chicken SHP2 (phospho-Tyr 542 ), anti-chicken JAK2, and anti-chicken TYK2 antibody (Biorbyt, San Francisco, CA, USA); rabbit anti-chicken GAPDH antibody (Abcam, Cambridge, MA, USA); Alexa Fluor® 488 Goat anti-rabbit IgG (H+L) secondary antibody (Invitrogen, Carlsbad, CA, USA); mouse monoclonal anti-chicken IFN-γ, IL-17A, IL-12p40 antibody and recombinants of these proteins (kindly provided by Dr. Hyun S. Lillehoj, USDA).

Techniques: Western Blot, Transfection, Quantitative RT-PCR

Journal: Frontiers in immunology

Article Title: PARP Inhibitor Upregulates PD-L1 Expression and Provides a New Combination Therapy in Pancreatic Cancer.

doi: 10.3389/fimmu.2021.762989

Figure Lengend Snippet: FIGURE 4 | Pamiparib treatment induces PD-L1 expression via JAK2/STAT3 pathway. (A) Cells were pretreated with pamiparib (100 mM, 12h) and PD-L1 expression was assessed by immunoblotting after treatment with the concentrations (20 mM) of AG490 for 24h. (B) Cells were pretreated with pamiparib (100 mM, 12h) and PD-L1 expression was assessed by protein blotting after treatment with stattic (20 mM) for 24h. (C) Protein expression of phospho-JAK2 (p-JAK2), JAK2, and PD-L1 in SW1990 cells after being treated with pamiparib (100 mM) for the indicated times. (D) Protein expression of phospho-STAT3(p-STAT3), STAT3 and PD-L1 in SW1990 cells after being treated with pamiparib (100 mM) for the indicated times. GAPDH was used as a loading control. (E) IHC staining of p-STAT3 of nude mouse xenograft tumors in control group and treated with pamiparib group. Scale bar, 100 mm. (F) IHC staining score showed the expression of p-STAT3 was significantly upregulated in nude mice pamiparib treated group. Data are mean ± SD; n = 3 samples per group. The IHC results were analyzed by Pearson c2 test. *P < 0.05.

Article Snippet: The primary antibodies are PD-L1 (CST #13684, Cell Signaling Technology), PD-L1 (17952- 1-AP, Santa Cruz), STAT3 (CST #9139, Cell Signaling Technology), phosphorylated STAT3 (CST #9145, Cell Signaling Technology), JAK2 (17670-1-AP, Proteintech), phosphorylated JAK2 (CST #4406, Cell Signaling Technology), AKT (10176-2-AP, Proteintech), phosphorylated AKT (66444-1-Ig, Proteintech), ERK (CST #4696, Cell Signaling Technology), phosphorylated ERK (CST #3510, Cell Signaling Technology), PARP-1 (sc-8007, Santa Cruz), and GAPDH (60004-1-Ig, Proteintech).

Techniques: Expressing, Western Blot, Control, Immunohistochemistry

Journal: Frontiers in immunology

Article Title: PARP Inhibitor Upregulates PD-L1 Expression and Provides a New Combination Therapy in Pancreatic Cancer.

doi: 10.3389/fimmu.2021.762989

Figure Lengend Snippet: FIGURE 8 | Diagram summarizing that pamiparib treatment induces PD-L1 expression mainly via JAK2/STAT3 in pancreatic cancer (details provided in the Discussion section).

Article Snippet: The primary antibodies are PD-L1 (CST #13684, Cell Signaling Technology), PD-L1 (17952- 1-AP, Santa Cruz), STAT3 (CST #9139, Cell Signaling Technology), phosphorylated STAT3 (CST #9145, Cell Signaling Technology), JAK2 (17670-1-AP, Proteintech), phosphorylated JAK2 (CST #4406, Cell Signaling Technology), AKT (10176-2-AP, Proteintech), phosphorylated AKT (66444-1-Ig, Proteintech), ERK (CST #4696, Cell Signaling Technology), phosphorylated ERK (CST #3510, Cell Signaling Technology), PARP-1 (sc-8007, Santa Cruz), and GAPDH (60004-1-Ig, Proteintech).

Techniques: Expressing