Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 5. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in medulloblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in medulloblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 6. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in glioblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in glioblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: PLoS ONE

Article Title: A Jak1/2 inhibitor, baricitinib, inhibits osteoclastogenesis by suppressing RANKL expression in osteoblasts in vitro

doi: 10.1371/journal.pone.0181126

Figure Lengend Snippet: (A) Immunoblot images of adenovirus-mediated knockdown of murine Jak1 or Jak2. (B, C) Effects of shJak1_#1, #2, shJak2_#1 or #2 on osteoclast formation in co-cultures of osteoblasts and bone marrow cells treated with 1,25D 3 and PGE 2 (B) or in bone marrow macrophage cultures treated with RANKL and M-CSF (C) . (D, E) Effects of shJak1_#1, #2, shJak2_#1 or #2 on expression of RANKL mRNAs (D) and proteins (E) in osteoblasts. Osteoblasts were transfected with shCtrl (lane 1, lane 2), shJak1_#1 (lane 3), shJak1_#2 (lane 4), shJak2_#1 (lane 5) or shJak2_#2 (lane 6) and treated with (lane 2–6) or without (lane 1) 1,25D 3 and PGE 2 for 24 h. In B–D : error bars, s.e. (n = 3–4). ** P < 0.01, Student's t test. † † P < 0.01, Dunnett’s multiple comparisons test (vs shCtrl with 1,25D 3 and PGE 2 ). Original immunoblot images are shown in .

Article Snippet: Immunoblotting was performed using the following antibodies; anti-phospho Jak1 (Tyr1022/1023) rabbit IgG (3331; Cell Signaling Technology, Beverly, MA, 1:1000), anti-Jak1 rabbit IgG (3332; Cell Signaling Technology, 1:1000), anti-phospho Jak2 (Tyr1007/1008) rabbit IgG (3776; Cell Signaling Technology, 1:1000), anti-Jak2 rabbit IgG (3230; Cell Signaling Technology, 1:1000), anti-RANKL goat IgG (sc-7628; Santa Cruz Biotechnology, Santa Cruz, CA, 1:1000), anti-phospho Stat3 (Tyr705) rabbit IgG (9145; Cell Signaling Technology, 1:2000), anti-Stat3α rabbit IgG (8768; Cell Signaling Technology, 1:1000), anti-α tubulin mouse IgG (CP06; Calbiochem, San Diego, CA, 1:1000), donkey anti-rabbit IgG-HRP (NA934V; GE Healthcare, Little Chalfont, UK, 1:5000), goat anti-mouse IgG-HRP (170–6516; Bio-Rad Laboratories, Hercules, CA, 1:2000), and donkey anti-goat IgG-HRP (sc-2056; Santa Cruz Biotechnology, 1:5000).

Techniques: Western Blot, Knockdown, Expressing, Transfection

Journal: PLoS ONE

Article Title: A Jak1/2 inhibitor, baricitinib, inhibits osteoclastogenesis by suppressing RANKL expression in osteoblasts in vitro

doi: 10.1371/journal.pone.0181126

Figure Lengend Snippet: (A) Densitometric data of cytokine protein array in co-cultured medium of osteoblasts and bone marrow cells in the presence or absence of 1,25D 3 and PGE 2 . Two independent experiments were performed, and a representative result is shown. (B, C) Osteoblasts were stimulated by 1,25D 3 and PGE 2 for 0.5 (B) or 1 h (B, C) , and phosphorylation of Jak1, Jak2, and Stat3 were determined by immunoblotting. (D) Effects of 2.5 μM baricitinib on expression of Socs3 mRNA in osteoblasts in the presence of 1,25D 3 and PGE 2 . (E) An activator of Stat3, colivelin (Santa Cruz Biotechnology; 0.1 or 1 µM), rescued baricitinib-induced RANKL down-regulation in osteoblast cultures. error bars, s.e. (n = 3). ** P < 0.01, Student’s t test. Original immunoblot images are shown in .

Article Snippet: Immunoblotting was performed using the following antibodies; anti-phospho Jak1 (Tyr1022/1023) rabbit IgG (3331; Cell Signaling Technology, Beverly, MA, 1:1000), anti-Jak1 rabbit IgG (3332; Cell Signaling Technology, 1:1000), anti-phospho Jak2 (Tyr1007/1008) rabbit IgG (3776; Cell Signaling Technology, 1:1000), anti-Jak2 rabbit IgG (3230; Cell Signaling Technology, 1:1000), anti-RANKL goat IgG (sc-7628; Santa Cruz Biotechnology, Santa Cruz, CA, 1:1000), anti-phospho Stat3 (Tyr705) rabbit IgG (9145; Cell Signaling Technology, 1:2000), anti-Stat3α rabbit IgG (8768; Cell Signaling Technology, 1:1000), anti-α tubulin mouse IgG (CP06; Calbiochem, San Diego, CA, 1:1000), donkey anti-rabbit IgG-HRP (NA934V; GE Healthcare, Little Chalfont, UK, 1:5000), goat anti-mouse IgG-HRP (170–6516; Bio-Rad Laboratories, Hercules, CA, 1:2000), and donkey anti-goat IgG-HRP (sc-2056; Santa Cruz Biotechnology, 1:5000).

Techniques: Protein Array, Cell Culture, Phospho-proteomics, Western Blot, Expressing

Journal: bioRxiv

Article Title: Spatial mapping of proteins and their activity states in cancer models by multiplex in situ PLA

doi: 10.1101/2025.07.11.662357

Figure Lengend Snippet: misPLA analyses of phosphorylation states and protein-protein interactions in SK-BR cells with or without EGF stimulation. A) Phosphorylation targets: SK-BR cells were analyzed for phosphorylation of STAT5a (pSTAT5a), STAT3 (pSTAT3), AKT (pAKT), ERK (pERK), and EGFR (pEGFR), under unstimulated and EGF-stimulated conditions. All targets were visualized three at a time in sequential detection cycles and are shown simultaneously (upper panels, “All targets”) and subsequently as individual channels. PLA signals (red, cyan, green, purple) reflect activated protein states detected via dual-recognition proximity ligation. DAPI (blue) labels nuclei. Scale bars = 50 µm. B) Protein-protein interactions: Visualization of the following protein-protein interactions investigated by misPLA in unstimulated and EGF-stimulated SK-BR cells: MEK1–ERK2, GRB2–MEK1, EGFR–GRB2, STAT3–STAT5a, JAK1–JAK3, JAK1–PI3K, JAK2–STAT5a, JAK1–STAT3, and JAK2–JAK3. Upper panels (“All targets”) represent simultaneous visualization of all targets, imaged three at a time in sequential detection cycles, followed by separated signals per interaction. Scale bars = 50 µm.

Article Snippet: The following primary antibodies were used for Western blotting: JAK1 (ProteinTech, 66466-1-Ig), STAT3 (ProteinTech, 60199-1-Ig; Abcam, ab171359), MEK1 (Abcam, ab239802), EGFR (Abcam, ab271834), AKT2 (Thermo Scientific, PA5-85518), ERK2 (Thermo Fisher, PA5-29636), phospho-PI3K p85/p55 (Cell Signaling Technology, 4228S), pSTAT3-Y705 (R&D Systems, AF4607), Grb2 (R&D Systems, mab38461), GAPDH (CST, 14C10), and Vinculin (CST, E1E9V), used at either 1:1000 or 1:2000 dilution.

Techniques: Phospho-proteomics, Protein-Protein interactions, Ligation

Journal: bioRxiv

Article Title: Spatial mapping of proteins and their activity states in cancer models by multiplex in situ PLA

doi: 10.1101/2025.07.11.662357

Figure Lengend Snippet: misPLA mapping of signaling interactions in a lymph-node Hodgkin lymphoma, mixed cellularity (right neck); Hodgkin lymphoma, lymphocyte-depleted (neck); Hodgkin lymphoma, lymphocyte-predominant (left neck); Hodgkin lymphoma, mixed cellularity (left neck); and thymoma type B3 (mediastinum). Top row (visualization cycle 1) displays MEK1–ERK2 (FITC), EGFR–GRB2 (Cy5) and GRB2–MEK1 (Cy3N) together with DAPI. Middle row (cycle 2) shows STAT3–STAT5a (FITC), JAK1–JAK3 (Cy5) and JAK1–PI3Kp85 (Cy3N). Bottom row (cycle 3) presents JAK2– STAT5a (FITC), JAK2–JAK3 (Cy5) and JAK1–STAT3 (Cy3N). All nine pairs of antibody-oligonucleotide conjugates were applied and then amplified in a single incubation. The RCA products were revealed using detection oligonucleotides conjugated with three fluorophores in three visualization cycles. A standard three-channel fluorescence microscope was used with identical settings for all three fluorophores. Scale bars, 50 µm.

Article Snippet: The following primary antibodies were used for Western blotting: JAK1 (ProteinTech, 66466-1-Ig), STAT3 (ProteinTech, 60199-1-Ig; Abcam, ab171359), MEK1 (Abcam, ab239802), EGFR (Abcam, ab271834), AKT2 (Thermo Scientific, PA5-85518), ERK2 (Thermo Fisher, PA5-29636), phospho-PI3K p85/p55 (Cell Signaling Technology, 4228S), pSTAT3-Y705 (R&D Systems, AF4607), Grb2 (R&D Systems, mab38461), GAPDH (CST, 14C10), and Vinculin (CST, E1E9V), used at either 1:1000 or 1:2000 dilution.

Techniques: Amplification, Incubation, Fluorescence, Microscopy

Journal: Naunyn-Schmiedeberg's archives of pharmacology

Article Title: Modulation of AMPK by esomeprazole and canagliflozin mitigates methotrexate-induced hepatotoxicity: involvement of MAPK/JNK/ERK, JAK1/STAT3, and PI3K/Akt signaling pathways.

doi: 10.1007/s00210-025-03908-3

Figure Lengend Snippet: Fig. 7 The regulation of the p-JAK1/p-STAT3 signaling pathway by ESOM and CANA. To verify the effect of ESOM and CANA on various groups, we measured A the relative protein expression of p-JAK1 via western blot analysis and B the relative protein expres- sion of p-STAT3 using western blot analysis. Data are shown as mean ± SE (n = 6). *, **, # indicate a significant difference between the control group, MTX group, and ESOM and CANA + MTX group, respec- tively, using one-way ANOVA test at a p value < 0.01 followed by Tukey

Article Snippet: The membranes were incubated at 4 °C overnight with 1:1000 dilutions of JAK1 antibody (Santa Cruz Biotechnology, sc-376996, USA), STAT3 antibody (Abcam Co., EPR787Y, UK), JNK antibody (Santa Cruz Biotechnology, sc-7345, USA), ERK1 antibody (Santa Cruz Biotechnology, sc-271270, USA), and p38 antibody (Biorbyt, orb127559, USA), respectively, then incubated at room temperature for 1 h with 1:2000 dilutions of HRP-linked anti-rabbit antibody (Cell Signaling Technology, 7074, USA).

Techniques: Expressing, Western Blot, Control

Journal: Scientific Reports

Article Title: 3-oxo-C12:2-HSL, quorum sensing molecule from human intestinal microbiota, inhibits pro-inflammatory pathways in immune cells via bitter taste receptors

doi: 10.1038/s41598-022-13451-3

Figure Lengend Snippet: 3-oxo-C12:2-HSL prevents activation of the JAK-STAT pathway. Cells were stimulated with LPS and IFNγ in absence (control) or in presence of 50 µM 3-oxo-C12:2-HSL for 2 h. The levels of phosphorylated proteins P-JAK1 ( a ), STAT1 ( b ), JAK2 ( c ), STAT3 ( d ) were normalized to their respective unphosphorylated forms. Results are expressed as mean ± SEM from 4 independent experiments. One-Way ANOVA, Dunnett’s post-test **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. control. ( e ) Reconstructed images from Simple Western analysis of protein levels and actin used as housekeeping protein are displayed. They are based on the area under the chemiluminescence signal curve obtained for one experiment, representative of 4 independent experiments performed in duplicates. Molecular markers are indicated on the left (kDa). 3oC12:2 stands for 3-oxo-C12:2-HSL.

Article Snippet: The microplates were loaded with 0.8 μg/μL protein, primary antibodies (as detailed below), and reagents provided by the manufacturer: anti-JAK1 (1:25; Biotechne MAB4260), anti-Phospho-JAK1 (1:100; Invitrogen 44422G), anti-JAK2 (1:13; Cell Signalling, 3230), anti-Phospho-JAK2 (1:13; Cell Signalling 3776), anti-STAT1 (1:50; Cell Signalling 9172), anti-Phospho-STAT1 (1:40; Biotechne AF2894), anti-Actin (1:1000; Millipore MAB1501R).

Techniques: Activation Assay, Western Blot

Journal: Scientific Reports

Article Title: 3-oxo-C12:2-HSL, quorum sensing molecule from human intestinal microbiota, inhibits pro-inflammatory pathways in immune cells via bitter taste receptors

doi: 10.1038/s41598-022-13451-3

Figure Lengend Snippet: Proposed mechanisms of 3-oxo-C12:2-HSL effects on inflammation. In immune cells, 3-oxo-C12:2-HSL is able to activate bitter taste receptors, which are G-protein coupled receptors, triggering a signalling cascade resulting in the release of calcium from the endoplasmic reticulum. In addition, in murine activated macrophages the presence of 3-oxo-C12:2-HSL attenuates the activation of the JAK-STAT signalling pathways, by specifically preventing JAK1 and STAT1 phosphorylation. This leads to a decrease in pro-inflammatory cytokine secretions and an overall reduced inflammatory response. Part of the effects of 3-oxo-C12:2-HSL on inflammatory response is dependent on bitter taste receptors. Created with BioRender.com.

Article Snippet: The microplates were loaded with 0.8 μg/μL protein, primary antibodies (as detailed below), and reagents provided by the manufacturer: anti-JAK1 (1:25; Biotechne MAB4260), anti-Phospho-JAK1 (1:100; Invitrogen 44422G), anti-JAK2 (1:13; Cell Signalling, 3230), anti-Phospho-JAK2 (1:13; Cell Signalling 3776), anti-STAT1 (1:50; Cell Signalling 9172), anti-Phospho-STAT1 (1:40; Biotechne AF2894), anti-Actin (1:1000; Millipore MAB1501R).

Techniques: Activation Assay