Figure 2 : Induction of apoptosis in leukemic cells by CASP3*-LENA.

From: Therapeutic potential of HIV protease-activable CASP3

Figure 2

(a) Enhancement of WT- and CASP3*-LENA production by a low dose of saquinavir (SQV, 0.2 µM). DMSO was used as a solvent control. The arrowhead indicates a signal representing the MA-CA, indicating the release of CASP3* from CASP3*-Gag. (b) Quantitative analysis of MOLT-4 cell viability after LENA exposure. Cell viability was scored by colorimetric assay at 1d post-LENA exposure comparing WT-LENA encapsidated with VSV-G (WT/VSV-G), non-enveloped CASP3*-LENA (ENV(-)), and CASP3*-LENA encapsidated with VSV-G. The average and standard deviation from triplicated wells using LENA produced in the presence or absence of SQV are shown (SQV and DMSO, respectively). (c) Light microscopic observation of MOLT-4 cells treated with LENA produced in the presence of low-dose SQV. Cells were imaged at 1 d post-LENA exposure. Cells were treated with either Bafilomycin A1 (BAF) or the CASP3 inhibitor Z-DEVD-FMK. DMSO was used as a solvent control. Bar, 50 µm; magnification x100. (d) Nuclear morphology of MOLT-4 cells treated with LENA produced in the presence of low-dose SQV. Cells were imaged at 1 d post-LENA exposure after staining with Hoechst dye. Cells were treated with either BAF or Z-DEVD-FMK. DMSO was used as a solvent control. Bar, 10 µm; magnification x630. (e) Flow cytometric detection of apoptotic MOLT-4 cells. Cells were stained with Annexin V Alexa Fluor 488 at 6 h post-exposure to LENA produced in the presence of low dose nelfinavir (NFV, 0.2 µM). The live cell fraction was gated to analyze the early phase of apoptotic cell death. Cells were treated with either BAF or Z-DEVD-FMK. DMSO was used as a solvent control. RLU, relative light units. (f) Quantitative analysis of MOLT-4 cell viability after exposure to LENA produced in the presence of low-dose NFV (0.2 µM). Cell viability was scored at 1d post-LENA exposure. Cells were treated with either BAF or Z-DEVD-FMK. DMSO was used as a solvent control. (g) Quantitative analysis of M8166 and SUP-T1 cell viabilities as perfomed on MOLT-4 cells.