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

Journal: Cancers

Article Title: Proteome-Wide Analysis Reveals TFEB Targets for Establishment of a Prognostic Signature to Predict Clinical Outcomes of Colorectal Cancer.

doi: 10.3390/cancers15030744

Figure Lengend Snippet: Figure 1. Mutations in S138 and S142 caused almost complete nuclear retention of TFEB. (A) Schematic of the human TFEB protein domains. S138 and S142 are localized in the proxim- ity of a NES. (B) S138 and S142 of TFEB are evolutionarily conserved in the indicated species. Alignment of the sequences near TFEB S138 and S142 is shown. (C) Western blots of endogenous and exogenous TFEB in CRC cells transfected with wild-type TFEB-GFP or TFEB-GFP mutant (S142A/S138A). Uncropped immunoblots are provided in the Figure S1. (D,E) Localization of TFEBWT and TFEBS142A/S138A in the cytosol and in the nucleus was detected by confocal microscopy. Representative images (D) and statistical results (E) are shown. Torin1 (250 nm), a positive control. Cell outlines in white dotted lines. DAPI was used to label the nucleus. Scale bars, 5 µm. Mean ± SD, n = 10 cells per condition, unpaired t-test. (F) Cell viability of indicated cells were analyzed using a CCK8 assay. Mean ± SEM, n = 3, unpaired t-test. ** p < 0.01, *** p < 0.001.

Article Snippet: The primary antibodies (against TFEB, GFP, LAMP2, NDRG1, SMPD1, and actin) and the HRP-conjugated secondary antibodies (goat anti-rabbit antibody) were supplied by Proteintech (Wuhan, China).

Techniques: Western Blot, Transfection, Mutagenesis, Confocal Microscopy, Positive Control, CCK-8 Assay

Journal: Longevity & Healthspan

Article Title: The secreted protein S100A7 (psoriasin) is induced by telomere dysfunction in human keratinocytes independently of a DNA damage response and cell cycle regulators

doi: 10.1186/2046-2395-3-8

Figure Lengend Snippet: Colony-forming efficiency analysis of NHEK transduced with p14 ARF , p16 INK4A , p53 and TRF2 ΔBΔM . NHEKs were transduced with amphotropic retroviral particles using spinfection and, 48 h later, trypsinised and seeded at clonal density (7 × 10 3 cells per 6-well plate). Cells were cultured for 2 weeks under drug selection and finally fixed and stained with Rhodamine B to reveal keratinocyte colonies. Colony-forming efficiency, displayed as percentage and relative to the respective EV control, was calculated by dividing the total number of colonies obtained per well by the total number of cells seeded per plate (7,000). Photos show wells representative of the results obtained for each construct. Legend: GFP, empty vector control for p14 ARF , p16 INK4A and p53 ; EV, empty vector control for TRF2 ΔBΔM (DN). This is the result of a single experiment.

Article Snippet: Retroviral vectors pLPC-N MYC (12540 Addgene, Cambridge, MA.) and pLPC-NMYC TRF2 ΔBΔM (16069 Addgene) were donated by Titia de Lange (Rockefeller University, NYC, USA); pBABE-puro p14 ARF , pBABE-puro p16 INK4a and pBABE-puro p53 were donated by Gordon Peters (London Research Institute, CRUK, London, UK); and pBABE-puro GFP was donated by Cleo Bishop (Blizard Institute, QMUL, London, UK).

Techniques: Transduction, Retroviral, Cell Culture, Selection, Staining, Control, Construct, Plasmid Preparation