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. 2012 Apr 30;197(3):361–367. doi: 10.1083/jcb.201111141

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© 2012 Huang et al.

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Figure 5. — Higher Bax activation in Cdh1 knockdown cells as a result of MOAP-1. (A) 293T cells treated with Cdh1 (siCdh1), MOAP-1 (siMOAP-1), or control siRNA (siCtr) were treated with control or 100 µM etoposide for 48 h. Bax activation was monitored by IP with 6a7 antibody from 293T cells followed by immunoblotting of total Bax (N20). Input lysates were immunoblotted with anti-actin and -Bax (N20) antibodies. (B) Cell lysates from A were immunoblotted with anti-Cdh1 or anti–MOAP-1 antibodies. (C) Control, Cdh1, and MOAP-1 knockdown PC3 cells were treated with 100 µM etoposide for 48 h. The percentage of cells with sub-G1 DNA content was measured by PI staining and flow cytometry. (D) PC3 cells were processed as in C but treated with 20 µM cisplatin. (C and D) The asterisks indicate that the difference between the two experiments is significant (P < 0.05), using an unpaired t test. The data shown represent three independent experiments. Error bars are SEM. (E) Cdh1 or MOAP-1 was knocked down using siRNA in PC3 cells, and samples were immunoblotted for Cdh1, MOAP-1, or actin. (F) Cells treated as in D were lysed, and lysates were immunoblotted to detect caspase 3 processing.