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. Author manuscript; available in PMC: 2022 Nov 4.

Published in final edited form as: Mol Cancer Res. 2022 May 4;20(5):699–711. doi: 10.1158/1541-7786.MCR-21-0837

Figure 2. — Knockdown of Pak2 diminishes metastatic colonization of Nf2-deficient mesothelioma cells in the lung. Cell line MM87, which was derived from asbestos-induced mesothelioma from a Nf2^+/−;Cdkn2a^+/− mouse, was infected with tet-inducible lentiviruses against Pak2. (A) Immunoblot demonstrating expression of Pak2 after knockdown with shRNA. Two clones with robust knockdown of Pak2 (shPak2 #70 and shPak2 #85) and one clone infected with lentivirus against GFP, used as a control, were selected for tail vein injections into NSG mice. (B) Malignant mesothelioma clones were each injected into the tail vein of three different NGS mice, followed by injection with doxycycline 7 d later and every 2 d thereafter; all animals were sacrificed on d 21, and lungs were collected for histopathological assessment of tumor burden. (C) H&E staining illustrating representative tumor colonization (intensely stained areas) of lungs by MM87 cells expressing shGFP, shPak2 #70 or shPak2 #85. Tumor burden was quantified using an Aperio ScanScope CS2 scanner, as described in the Supplemental Materials and Methods.

Figure 2. — Knockdown of Pak2 diminishes metastatic colonization of Nf2-deficient mesothelioma cells in the lung. Cell line MM87, which was derived from asbestos-induced mesothelioma from a Nf2^+/−;Cdkn2a^+/− mouse, was infected with tet-inducible lentiviruses against Pak2. (A) Immunoblot demonstrating expression of Pak2 after knockdown with shRNA. Two clones with robust knockdown of Pak2 (shPak2 #70 and shPak2 #85) and one clone infected with lentivirus against GFP, used as a control, were selected for tail vein injections into NSG mice. (B) Malignant mesothelioma clones were each injected into the tail vein of three different NGS mice, followed by injection with doxycycline 7 d later and every 2 d thereafter; all animals were sacrificed on d 21, and lungs were collected for histopathological assessment of tumor burden. (C) H&E staining illustrating representative tumor colonization (intensely stained areas) of lungs by MM87 cells expressing shGFP, shPak2 #70 or shPak2 #85. Tumor burden was quantified using an Aperio ScanScope CS2 scanner, as described in the Supplemental Materials and Methods.