mouse nras  (Santa Cruz Biotechnology)

Journal: Aging Cell

Article Title: Loss of p62 Binding Allows TIF ‐ IA Accumulation in Senescence, Which Promotes Phenotypic Changes to Nucleoli and the Senescence Associated Secretory Phenotype

doi: 10.1111/acel.70334

Figure Lengend Snippet: TIF‐IA protein accumulates in an animal model of OIS and in chronological and genetic models of ageing. (A) TIF‐IA and N‐ras immunohistochemistry were performed on liver sections from mice culled 6 days after hydrodynamic delivery of Nras G12V/D38A ( n = 5) or Nras G12V ( n = 5) transposons. Left: Representative images at 10× (scale bar 100 μm) and 40× (scale bar 20 μm) magnification. Right: QuPath software (v0.2.3) was used to quantify the percentage of TIF‐IA and N‐ras positive cells in five regions of interest (ROIs) per section. Pooled data (25 ROIs per condition) is shown. Mann–Whitney test determined significance. (B) Anti‐TIF‐IA immunohistochemistry was performed on colonic sections from young and old wild type (WT) and nfkb1 −/− mice ( n = 5 mice per condition). Left: Representative images at 10× and 40× magnification. Scale bar 20 μm. Right: QuPath software (v0.2.3) was used to quantify nuclear TIF‐IA intensity in five ROIs as above. Pooled data are shown. A Kruskal–Wallis test with Dunns multiple comparison was used to determine significance. ns, nonsignificant.

Article Snippet: Sections were incubated overnight at 4°C with the primary antibodies: rabbit RRN3 (Abcam 251933) and mouse NRas (Santa Cruz Biotechnology sc‐31), diluted 1:500 in antibody diluent (Dako).

Techniques: Animal Model, Immunohistochemistry, Software, MANN-WHITNEY, Comparison

Journal: EMBO Reports

Article Title: Proximity-labeling proteomics reveals remodeled interactomes and altered localization of pathogenic SHP2 variants

doi: 10.1038/s44319-025-00674-4

Figure Lengend Snippet: ( A ) In vitro activity measurements showing that SHP2-TurboID can be activated by a phosphopeptide (PD-1 pY248) ( n = 3). Bar heights indicate the mean, and error bars indicate standard deviation. ( B ) In vitro activity measurements showing that E76K hyperactivation is preserved in a TurboID-fusion context, and only SHP2 WT -TurboID, but not SHP2 E76K -TurboID can be further activated by PD-1 pY248 ( n = 3). Bar heights indicate the mean, and error bars indicate standard deviation. ( C ) Schematic of the Ras dephosphorylation assay in HEK 293 cells. ( D ) Representative western blots showing dephosphorylation of N-Ras by SHP2-TurboID proteins. WT = SHP2 WT EK = SHP2 E76K . ( E ) Quantification of Ras dephosphorylation assays ( n = 4). Bar heights indicate the mean, and error bars indicate standard deviation. ( F ) The left blot compares SHP2 levels in WT and SHP2 KO HEK 293 cells, and shows over-expression of SHP2 WT -TurboID in SHP2 KO cells, highlighting partial cleavage of SHP2-TurboID. The right blot compares endogenous SHP2 levels in WT HEK 293 cells with SHP2 WT -TurboID levels in SHP2 KO HEK 293 cells, transfected under the same conditions used for proteomics experiments. ( G ) Further confirmation of SHP2 KO cells used for proteomics experiments, comparing SHP2 levels detected by mass spectrometry in negative control TurboID-only samples and SHP2 WT -TurboID samples. Bar heights indicate the mean, and error bars indicate standard deviation. ( H ) Fold-change between SHP2 WT -TurboID and TurboID-only for total lysates, measuring protein abundance (x-axis), and streptavidin-IP, measuring proximity labeling (y-axis), in unstimulated cells. ( I ) Same as ( H ), but for cells stimulated with 100 ng/mL EGF ( n = 3 for total lysate and TurboID datasets). .

Article Snippet: pCGN N-Ras wt , Addgene , #14723.

Techniques: In Vitro, Activity Assay, Phospho-proteomics, Standard Deviation, De-Phosphorylation Assay, Western Blot, Over Expression, Transfection, Mass Spectrometry, Negative Control, Quantitative Proteomics, Labeling