Inhibition of Hsp90 Leads to Cell Cycle Arrest and Apoptosis in Human Malignant Pleural Mesothelioma Junichi Okamoto, MD, PhD, Iwao Mikami, MD, PhD,

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Inhibition of Hsp90 Leads to Cell Cycle Arrest and Apoptosis in Human Malignant Pleural Mesothelioma Junichi Okamoto, MD, PhD, Iwao Mikami, MD, PhD, Yuichi Tominaga, PhD, Kristopher M. Kuchenbecker, BA, Yu-Ching Lin, MD, Dawn T. Bravo, PhD, Genevieve Clement, PhD, Adam Yagui-Beltran, MD, M Roshni Ray, BS, Kiyoshi Koizumi, MD, Biao He, PhD, David M. Jablons, MD Journal of Thoracic Oncology Volume 3, Issue 10, Pages 1089-1095 (October 2008) DOI: 10.1097/JTO.0b013e3181839693 Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Western analysis of the Hsp90 expression in 13 MM tissue samples (A) and in 5 MM cell lines (B). β-Actin served as loading control. Journal of Thoracic Oncology 2008 3, 1089-1095DOI: (10.1097/JTO.0b013e3181839693) Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Cell cycle analysis of MM cells after 17-AAG treatments. (A) 17-AAG induces G1 arrest in MM cell lines. (B) 17-AAG induces G2/M cell cycle arrest in MM cell lines. All cell lines were treated with DMSO, 1 μm and 2 μm 17-AAG for 24 hours. Journal of Thoracic Oncology 2008 3, 1089-1095DOI: (10.1097/JTO.0b013e3181839693) Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Proliferation assay of MM cells after 17-AAG treatments. (A) MTT assay after 48 hours treatment with 17-AAG. A 17-AAG inhibits growth of MM cells. Gray: OD490 (DMSO treated cells)/OD490 (nontreated cells); black: OD490 (17-AAG treated cells)/OD490 (nontreated cells). (B) Western analysis of Hsp90, Hsp70, AKT, AKT1, and Survivin expression level in MM cell lines. β-Actin served as loading control. Journal of Thoracic Oncology 2008 3, 1089-1095DOI: (10.1097/JTO.0b013e3181839693) Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 A 17-AAG induces apoptosis in MM cell lines. (A) FACS analysis of apoptosis after17-AAG treatment in MM cell lines. Two days treatment: H28, REN, and H290. Three days treatment: H2052 and 211H. (B) Western analysis of expression levels of cleaved PARP and Survivin in MM cell lines. β-Actin served as loading control. Journal of Thoracic Oncology 2008 3, 1089-1095DOI: (10.1097/JTO.0b013e3181839693) Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 5 A 17-AAG induces apoptosis in MM primary tissue culture. (A) FACS analysis of apoptosis after 17-AAG treatment in MM primary culture. (B) Western analysis after 17-AAG treatment in MM primary culture. β-Actin served as loading control. Journal of Thoracic Oncology 2008 3, 1089-1095DOI: (10.1097/JTO.0b013e3181839693) Copyright © 2008 International Association for the Study of Lung Cancer Terms and Conditions