The Expression of AIP-Related Molecules in Elucidation of Cellular Pathways in Pituitary Adenomas Elina Heliövaara, Anniina Raitila, Virpi Launonen,

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The Expression of AIP-Related Molecules in Elucidation of Cellular Pathways in Pituitary Adenomas Elina Heliövaara, Anniina Raitila, Virpi Launonen, Anders Paetau, Johanna Arola, Heli Lehtonen, Timo Sane, Robert J. Weil, Outi Vierimaa, Pasi Salmela, Karoliina Tuppurainen, Markus Mäkinen, Lauri A. Aaltonen, Auli Karhu The American Journal of Pathology Volume 175, Issue 6, Pages 2501-2507 (December 2009) DOI: 10.2353/ajpath.2009.081131 Copyright © 2009 American Society for Investigative Pathology Terms and Conditions

Figure 1 ARNT and AHR immunohistochemistry in AIPmut+ and AIPmut- pituitary tumors at an original magnification of ×356. A: Negative ARNT staining in a patient with AIP Q14X mutation (left); positive ARNT staining in AIPmut− adenoma (right). B: Positive nuclear AHR staining in a patient with AIP Q14X mutation (left); negative nuclear AHR staining in AIPmut− adenoma (right). The American Journal of Pathology 2009 175, 2501-2507DOI: (10.2353/ajpath.2009.081131) Copyright © 2009 American Society for Investigative Pathology Terms and Conditions

Figure 2 Relative expressions of Aip in Aip+/+, Aip+/−, and Aip−/− MEF cell lines. SD is shown by error bars. The American Journal of Pathology 2009 175, 2501-2507DOI: (10.2353/ajpath.2009.081131) Copyright © 2009 American Society for Investigative Pathology Terms and Conditions

Figure 3 Western blot results from MEF, HeLa, and HEK293 cell lines. (Arrows indicate the bands corresponding to the proteins detected.) A: Arnt, Aip and α-Tubulin expression in Aip wild-type (Aip+/+) and Aip-null (Aip−/−) MEF cell lines. B: ARNT, AIP, and α-tubulin expression in HeLa and HEK293 cell lines: 1) untreated; 2) AIP siRNA-transfected; 3) control siRNA-transfected; and 4) transfection reagent-treated (DF1) cells. The American Journal of Pathology 2009 175, 2501-2507DOI: (10.2353/ajpath.2009.081131) Copyright © 2009 American Society for Investigative Pathology Terms and Conditions

Figure 4 Western blot and proliferation assay results from GH3 cell line. A: Arnt, Aip, and α-tubulin expression in 1) Aip siRNA transfected; 2) untreated; and 3) control siRNA-transfected cells. In each lane the intensity of Arnt protein has been normalized against α-tubulin. B: Cell proliferation of Aip siRNA, control siRNA-transfected, and untreated cells performed by MTS assay at different time points. The American Journal of Pathology 2009 175, 2501-2507DOI: (10.2353/ajpath.2009.081131) Copyright © 2009 American Society for Investigative Pathology Terms and Conditions