IHC Analysis of Ki67 and RFP

Immunohistochemistry (IHC) was performed as before[22 (link)]. Ki67 (Abcam, #ab15580) and RFP (Rockland, #600–401-379) staining was performed by the institutional Molecular Cytology Core using a Discovery XT system (Ventana Medical Systems, Roche). Slides were either imaged on an Axio wide-field microscope (Zeiss) or scanned with MIRAX scan (Zeiss) and analyzed with Pannoramic Viewer. Ki67 was quantified by manually counting stained nuclei in the field size as indicated. N = 7 and 9 biological replicates for the V558Δ and V558Δ;V653A groups, respectively.

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Zhang J.Q., Bosbach B., Loo J.K., Vitiello G.A., Zeng S., Seifert A.M., Medina B.D., Param N.J., Maltbaek J.H., Rossi F., Antonescu C.R., Besmer P, & DeMatteo R.P. (2020). The V654A second-site KIT mutation increases tumor oncogenesis and STAT activation in a mouse model of gastrointestinal stromal tumor. Oncogene, 39(49), 7153-7165.

Publication 2020

Discovery XT system Axio wide-field microscope MIRAX scan

Biological Cytology Immunohistochemistry Microscope Scan

Corresponding Organization :

Other organizations : Memorial Sloan Kettering Cancer Center, Hospital of the University of Pennsylvania

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Variable analysis

independent variables

V558Δ
V558Δ;V653A

dependent variables

Ki67 staining
RFP staining

control variables

Field size for Ki67 quantification

controls

Positive control: None specified
Negative control: None specified

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