Quantitative Immunohistochemical Analysis of IGF Signaling

Immunohistochemical analysis of tissue microarray comprising lung biopsy specimens was performed as described previously5 (link)69 (link). In brief, IHC staining was performed as described above using primary antibodies against IGF1 (Santa Cruz; Cat. No. sc-9013), IGF2 (Millipore; Cat. No. 05-166), IGF-1R (Cell Signaling; Cat. No. 3027), pIGF-1R (Cell Signaling; Cat. No. 3024). The expression level of IGF1, IGF2, IGF-1R, and pIGF-1R was blindly analysed and quantified using a four-value intensity score (0, 1+, 2+ and 3+) and a percentage (0–100%) of the extent of reactivity. A final expression score was obtained by multiplying the intensity value by the extent of reactivity (range, 0–300).

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Boo H.J., Min H.Y., Jang H.J., Yun H.J., Smith J.K., Jin Q., Lee H.J., Liu D., Kweon H.S., Behrens C., Lee J.J., Wistuba I.I., Lee E., Hong W.K, & Lee H.Y. (2016). The tobacco-specific carcinogen-operated calcium channel promotes lung tumorigenesis via IGF2 exocytosis in lung epithelial cells. Nature Communications, 7, 12961.

Publication 2016

IGF1 IGF-1R pIGF-1R

Antibodies Biopsy Igf 1r Igf1 Igf2 Lung Tissue microarray analysis

Corresponding Organization : Seoul National University

Other organizations : The University of Texas MD Anderson Cancer Center, Korea Basic Science Institute

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Variable analysis

independent variables

Primary antibodies against IGF1 (Santa Cruz; Cat. No. sc-9013)
Primary antibodies against IGF2 (Millipore; Cat. No. 05-166)
Primary antibodies against IGF-1R (Cell Signaling; Cat. No. 3027)
Primary antibodies against pIGF-1R (Cell Signaling; Cat. No. 3024)

dependent variables

Expression level of IGF1
Expression level of IGF2
Expression level of IGF-1R
Expression level of pIGF-1R

control variables

Lung biopsy specimens
Immunohistochemical analysis of tissue microarray

controls

No positive or negative controls were explicitly mentioned in the provided information.

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