American Society of Hematology

Figure 4

$Figure 4. Effects of shRNA-mediated knockdown of HDAC1 on bortezomib-induced apoptosis in MM cells. (A) MM cell lines were transfected with either pLL3.7-sh-HDAC1 (sh-HDAC1) or sh-control vector. Whole-cell lysates were prepared from GFP+ cells collected using a FACSAria flow cytometer and subjected to immunoblotting. The signal intensities of each band were quantified, normalized to those of the corresponding GAPDH, and shown as relative values setting the sh-control to 1.0. (B) MM cell lines transfected with shRNA vectors were cultured in the absence or presence of 2nM bortezomib. After 48 hours, MM cells were harvested, stained with annexin-V/APC, and subjected to flow cytometric analysis. The y-axis shows the proportion of annexin-V positivity in the GFP+ fraction. The means ± SD (bars) of 3 independent experiments are shown. P values were calculated by 1-way ANOVA with the Student-Newman-Keuls multiple comparisons test. *P < .05 against the sh-control. (C) shRNA-transduced RPMI8226 cells were cultured in the absence or presence of 2nM bortezomib. After 48 hours, whole-cell lysates were prepared from GFP+ cells collected by the FACSAria flow cytometer, and subjected to immunoblotting. The signal intensities of each band were quantified, normalized to those of the corresponding GAPDH, and shown as relative values setting the untreated sh-control to 1.0.$

Effects of shRNA-mediated knockdown of HDAC1 on bortezomib-induced apoptosis in MM cells. (A) MM cell lines were transfected with either pLL3.7-sh-HDAC1 (sh-HDAC1) or sh-control vector. Whole-cell lysates were prepared from GFP⁺ cells collected using a FACSAria flow cytometer and subjected to immunoblotting. The signal intensities of each band were quantified, normalized to those of the corresponding GAPDH, and shown as relative values setting the sh-control to 1.0. (B) MM cell lines transfected with shRNA vectors were cultured in the absence or presence of 2nM bortezomib. After 48 hours, MM cells were harvested, stained with annexin-V/APC, and subjected to flow cytometric analysis. The y-axis shows the proportion of annexin-V positivity in the GFP⁺ fraction. The means ± SD (bars) of 3 independent experiments are shown. P values were calculated by 1-way ANOVA with the Student-Newman-Keuls multiple comparisons test. *P < .05 against the sh-control. (C) shRNA-transduced RPMI8226 cells were cultured in the absence or presence of 2nM bortezomib. After 48 hours, whole-cell lysates were prepared from GFP⁺ cells collected by the FACSAria flow cytometer, and subjected to immunoblotting. The signal intensities of each band were quantified, normalized to those of the corresponding GAPDH, and shown as relative values setting the untreated sh-control to 1.0.

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