Journal: Science advances

Article Title: N-MYC impairs innate immune signaling in high-grade serous ovarian carcinoma.

doi: 10.1126/sciadv.adj5428

Figure Lengend Snippet: Fig. 3. N-MYC represses IFN type I immune signaling. (A) qRT-PCR of IFN type I (IFNA2 and IFNB1) and III (IFNL1, IFNL2, and IFNL3) ligands in CaOV3 MYCN/GFP and GFP cells treated ± DOX (1 μg/ml) for 72 hours. P values were calculated using an unpaired two-tailed Student’s t test. (B) qRT-PCR of IFNAR1 and IFNLR1 (top) and ISGs (bottom) in CaOV3 wild-type cells transfected with scrambled negative control siRNA or siRNAs specific for IFNAR1 and IFNLR1 for 72 hours. (C) Immunoblot of P-STAT1, STAT1, P-STAT2, STAT2, IRF9, and α-tubulin levels in CaOV3 MYCN/GFP and CaOV3 GFP cells treated ± DOX for 24 and 72 hours. Data are representative of three independent experiments. Paired comparisons are shown in the same color for densitometry fold changes. (D) Immunoblot with the indicated antibodies in CaOV3 MYCN/GFP and GFP cells pretreated ± DOX for 72 hours and then treated with IFNB1 (10 ng/ml) for 2 hours. Nuclear and cytoplasmic fractions were prepared and subjected to Western blot. Data are representative of three independent experiments. (E) ChIP-qPCR analysis of IRF9 binding to the ISRE sequence of the ISG15 promoter in CaOV3 MYCN/GFP and GFP cells pretreated ± DOX for 72 hours and then treated with IFNB1 (50 ng/ml) for 30 min. P values were calculated using one-way analysis of variance (ANOVA) with Tukey posttest for pairwise comparison. (F) qRT-PCR of multiple ISGs in CaOV3 MYCN/GFP and GFP cells pretreated ± DOX (1 μg/ml) for 72 hours followed by IFNB1 pulse (10 ng/ml) for 2 hours and then 24-hour chase. P values were calculated using one-way ANOVA with Tukey posttest for pairwise comparison. Data in (A), (B), (E), and (F) are means ± SEM of n = 3 biological replicates. *P < 0.05; **P < 0.005; ***P < 0.001; ****P < 0.0001. Densitometry analysis was performed using ImageJ software.

Article Snippet: The following antibodies were used in this study: anti- Hu Fc receptor binding inhibitor (#50- 112- 9053, eBioscience), P- STAT1 (Tyr701) (#9167, Cell Signaling Technology), STAT1 (#14994, Cell Signaling Technology), P- STAT2 (Tyr690) (#4441, Cell Signaling Technology), STAT2 (#72604, Cell Signaling Technology), IRF9 (#76684, Cell Signaling Technology), α- tubulin (#3873, Cell Signaling Technology), DYKDDDDK Tag rabbit monoclonal antibody (mAb) (#14793, Cell Signaling Technology), DYKDDDDK Tag mouse mAb (#8146, Cell Signaling Technology), STING (#13647, Cell Signaling Technology), TATA box–binding protein (#8515, Cell Signaling Technology), CD3- PECy7 (#341111, BD Bioscience Technology), RIG- I (#3743, Cell Signaling Technology), MDA- 5 (#5321, Cell Signaling Technology), P- TBK1 (Ser172) (#5483, Cell Signaling Technology), TBK1 (#3504, Cell Signaling Technology), P- IRF3 (Ser396) (#4947, Cell Signaling Technology), N- MYC (#51705, Cell Signaling Technology), VDAC (#4661, Cell Signaling Technology), MAVS (#24930, Cell Signaling Technology), c- MYC (#5605, Cell Signaling Technology), L- MYC (#76266, Cell Signaling Technology), P- STING (Ser366) (#50907, Cell Signaling Technology), hemagglutinin tag (#3724, Cell Signaling Technology), Myc tag (#2278, Cell Signaling Technology), cGAS (#15102, Cell Signaling Technology), anti- rabbit immunoglobulin G (IgG) (H+L) (DyLight 800 4× polyethylene glycol conjugate) (#5151, Cell Signaling Technology), and anti- mouse IgG (H+L) (DyLight 680 conjugate) (#5470, Cell Signaling Technology).

Techniques: Quantitative RT-PCR, Two Tailed Test, Transfection, Negative Control, Western Blot, ChIP-qPCR, Binding Assay, Sequencing, Comparison, Software

Journal: eLife

Article Title: Multiple tumor suppressors regulate a HIF-dependent negative feedback loop via ISGF3 in human clear cell renal cancer

doi: 10.7554/eLife.37925

Figure Lengend Snippet:

Article Snippet: Antibody , STAT2 (rabbit polyclonal) , Bethyl A303-512A , , , , (1:1,000 for western blot, 1:25 for IHC).

Techniques: Western Blot, Recombinant

Journal: Molecular Metabolism

Article Title: Baricitinib counteracts metaflammation, thus protecting against diet-induced metabolic abnormalities in mice

doi: 10.1016/j.molmet.2020.101009

Figure Lengend Snippet: Baricitinib attenuates the HD-induced JAK-STAT pathway. Western blotting analysis for phosphorylation of Tyr 1007/1008 JAK1/2 in the skeletal muscle (A) and kidneys (B) and normalized to total JAK1/2 and for phosphorylation of Tyr 690 on STAT2 in the skeletal muscle (C) and kidneys (D) and normalized to total STAT2. All of the data are expressed as mean ± SEM for n = 6 per group. ∗p< 0.05 vs ND and •p< 0.05 vs HD.

Article Snippet: The antibodies used were rabbit anti-Tyr 1007/1008 JAK2 (#3776), rabbit anti-total JAK2 (#3230), rabbit anti-Tyr 690 STAT2 (Bioss Antibodies, bs-3428R), rabbit anti-total STAT2 (#72604), rabbit p21 (#2947), rabbit anti-Ser 307 IRS-1 (#2381), mouse anti-total IRS-1 (#3194), rabbit anti-Ser 473 AKT (#4060), rabbit anti-total AKT (#9272), rabbit anti-Ser 9 GSK-3β (#9332) and rabbit anti-total GSK-3β (9315).

Techniques: Western Blot