Journal: Cell

Article Title: LECT2, a Ligand for Tie1, Plays a Crucial Role in Liver Fibrogenesis.

doi: 10.1016/j.cell.2019.07.021

Figure Lengend Snippet: Figure 4. Direct Interactions between LECT2 and Tie1 (A) Schematic illustration of Tie1 and LECT2 constructs. (B) Exogenous Myc-tagged LECT2 and Flag-tagged Tie1 were immunoprecipitated. (C) Endogenous LECT2 and Tie1 were immunoprecipitated.

Article Snippet: The following primary antibodies were used for co-immunoprecipitation and western blotting: Anti-Flag antibody (1:1000, SigmaAldrich, F1804, RRID:AB_262044), anti-MYC antibody (1:1000, Proteintech, 66004-1-Ig), anti-His antibody (1:1000, proteintech, 66005-1-Ig, RRID:AB_11232599), anti-LECT2 antibody (1:1000, Abcam, ab196015), anti-Tie1 antibody (1:500, Invitrogen, PA527903, RRID:AB_2545379), anti-Tie1Phospho-Tyr1117 antibody (1:500, Abcam, AB12535), anti-Tie2 antibody (1 mg/ml, R&D, AF313, RRID:AB_355295), anti-Tie2-Phospho-Y992 antibody (0.5 mg/ml, R&D, AF2720, RRID:AB_442172), anti-p38 antibody (1:1000, Abcam, ab31828), anti-PhosphoY182-p38 antibody (1:2000, Abcam, ab47363) anti-Met antibody (1:1000, Cell Signaling Technology, 8198, RRID:AB_10858224), anti-Met-Phospho-Tyr1234/1235 antibody (1:1000, Cell Signaling Technology, 3077, RRID:AB_2315156), anti-VEGFR2 antibody (1:1000, Cell Signaling Technology, 2479, RRID:AB_2212507), anti-VEGFR2-PhosphoTyr1175 antibody (1:1000, Cell Signaling Technology, 2478, RRID:AB_331377), anti-VE-cadherin (1:2000, Abcam, ab33168), and anti-a-Tubulin antibody (1:1000, Beijing Ray Antibody Biotech, RM2007).

Techniques: Construct, Immunoprecipitation

Journal: Molecular cell

Article Title: The MYCN oncoprotein is an RNA-binding accessory factor of the nuclear exosome targeting complex.

doi: 10.1016/j.molcel.2024.04.007

Figure Lengend Snippet: Figure 4. Exosome function controls the dynamics of the MYCN/MAX/MXD network (A) Immunoblot of IMR-32 cells stably expressing an shRNA targeting EXOSC10 treated for 72 h with Dox or EtOH as control. ACTB was loading control. Blot is representative of n = 3 independent replicates. (B) Read distribution of MYCN ChIP-Rx, MAX, MNT, and SIN3A cleavage under targets & release using nuclease (CUT&RUN) at the CDK5RAP3 and PHB genes. Cells treated as in (A) (n = 3 for MYCN ChIP-Rx; n = 2 for all C&Rs).

Article Snippet: 3 mg of each, MAX (Proteintech), MNT (Thermo Fisher Scientific), or SIN3A (Novus Biologicals) antibody were added per sample and incubated with shaking (800 rpm) at 4 C overnight.

Techniques: Western Blot, Stable Transfection, Expressing, shRNA, Control

Journal: Nature communications

Article Title: The transcriptional co-repressor Runx1t1 is essential for MYCN-driven neuroblastoma tumorigenesis.

doi: 10.1038/s41467-024-49871-0

Figure Lengend Snippet: Fig. 2 | Runx1t1 loss reverses MYCN-mediated sustained hyperplasia and induces ganglia neurite extension. a The percentage neuroblast hyperplasia scored from homozygous Th-MYCN (+/+) mice or littermate mice lacking the MYCN transgene (−/−), with either wild-type (+/+) or heterozygous loss (+/−) of Runx1t1. Scoring of N = 3–8 independent mice was performed for each genotype and timepoint. All data points were N = 3, except for +/+, +/+ week 1 and week 2 (N = 4); +/+, +/−day 0 (N = 8) and week 4 (N = 4); −/−, +/−day 0 (N = 6), week 1 (N = 4) and week 4 (N = 5). The graph is mean ± SEM. b Representative histology of RUNX1T1 staining in ganglia from mice homozygous for the Th-MYCN transgene, and either wild-type or heterozygous for Runx1t1 from day 0 and 4 weeks of age.

Article Snippet: Samples were embedded in paraffin, sectioned, and stained with H&E or for MYCN (rabbit polyclonal antibody, 10159-2-AP, Proteintech, 1:1000) and RUNX1T1 (as described above).

Techniques: Staining