Figure 3
KLF9 knockdown suppresses apoptosis and efficient up-regulation of NOXA by bortezomib or LBH589. (A) MM1.S cells were infected with control shRNAs (CL), KLF9 shRNAs 1 (K9-1), or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the vehicle (DMSO) or with the indicated amounts of bortezomib (BTZ) or LBH589 (LBH) for 48 hours. Total protein extracts from the cells were probed by Western blotting with antibodies designated on the left. *Endogenous NOXA and cleaved caspase 3 levels in untreated cells were not detected under the exposure conditions used to detect these proteins in cells treated with the drugs. (B) Cells described in panel A were stained with the trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields. The 2-tailed P value is shown for the groups with close values of the means. (C) RPMI-8226 cells were infected with control shRNAs (CL) or KLF9 shRNAs 1 (K9-1) or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the vehicle (DMSO) or with the indicated amounts of bortezomib (BTZ) or LBH589 (LBH) for 48 hours. Total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. *Endogenous NOXA and cleaved caspase 3 levels in untreated cells were not detected under the exposure conditions used to detect these proteins in cells treated with the drugs. (D) Cells described in panel C were stained with trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields. The two-tailed P value is shown for the groups with close values of the means. (E) Cells were incubated with no drug (UT) or with the indicated amounts of bortezomib (BTZ), LBH589 (LBH), or combination of the same concentration of both drugs for 24 hours. After incubation, cells were collected and total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. (F) Cells were infected with control shRNAs (CL) KLF9 shRNAs 1 (K9-1), or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the indicated amounts of bortezomib (BTZ), LBH589 (LBH), or combination of the same concentration of both drugs. After incubation, cells were collected and total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. (G) Cells described in panel F were stained with trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields.

KLF9 knockdown suppresses apoptosis and efficient up-regulation of NOXA by bortezomib or LBH589. (A) MM1.S cells were infected with control shRNAs (CL), KLF9 shRNAs 1 (K9-1), or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the vehicle (DMSO) or with the indicated amounts of bortezomib (BTZ) or LBH589 (LBH) for 48 hours. Total protein extracts from the cells were probed by Western blotting with antibodies designated on the left. *Endogenous NOXA and cleaved caspase 3 levels in untreated cells were not detected under the exposure conditions used to detect these proteins in cells treated with the drugs. (B) Cells described in panel A were stained with the trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields. The 2-tailed P value is shown for the groups with close values of the means. (C) RPMI-8226 cells were infected with control shRNAs (CL) or KLF9 shRNAs 1 (K9-1) or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the vehicle (DMSO) or with the indicated amounts of bortezomib (BTZ) or LBH589 (LBH) for 48 hours. Total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. *Endogenous NOXA and cleaved caspase 3 levels in untreated cells were not detected under the exposure conditions used to detect these proteins in cells treated with the drugs. (D) Cells described in panel C were stained with trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields. The two-tailed P value is shown for the groups with close values of the means. (E) Cells were incubated with no drug (UT) or with the indicated amounts of bortezomib (BTZ), LBH589 (LBH), or combination of the same concentration of both drugs for 24 hours. After incubation, cells were collected and total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. (F) Cells were infected with control shRNAs (CL) KLF9 shRNAs 1 (K9-1), or KLF9 shRNAs2 (K9-2). Forty-eight hours after infection, cells were incubated with the indicated amounts of bortezomib (BTZ), LBH589 (LBH), or combination of the same concentration of both drugs. After incubation, cells were collected and total protein extracts from the cells were probed in Western blotting with antibodies designated on the left. (G) Cells described in panel F were stained with trypan blue. The percentage of dead cells was determined by counting positive and negative cells under light microscope in multiple view fields.

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