BD PharmingenTM MitoStatus TMRE

Description

Panel 1. Flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO solid line histograms , 5 muM camptothecin Left Plot, dashed line histogram for 4 hr or 50 muM FCCP 20 min, Right Plot, dashed line histogram , and then stained with 100 nM BD Pharmingen MitoStatus TMRE 15 min, Cat. No. 564696 . Panel 2. Two-color flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO Left Plot or 5 muM camptothecin Right Plot for 4 hr, then stained with 100 nM MitoStatus TMRE 15 min . Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with APC Annexin V 15 min, Cat. No. 556419 . Co-staining shows two main populations: healthy cells that are MitoStatus TMRE-positive and Annexin V-negative, and apoptotic or dead cells that are MitoStatus TMRE-negative and APC Annexin V-positive. Compared to the DMSO-vehicle treated control, camptothecin treatment increased the MitoStatus TMRE-negative and APC Annexin V-positive population, indicating that more cells are undergoing apoptosis. Loss of mitochondrial membrane potential is an early apoptotic marker so a small population of cells in transition is MitoStatus TMRE-negative and APC Annexin V-negative. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 3. Immunofluorescent imaging of TMRE in HeLa Cells. HeLa cells were treated with 0.02 % DMSO Left Image or 1 uM staurosporine Right Image for 3 hr. Cells were stained 30 min with 200 nM MitoStatus TMRE and 5 ug/mL Hoechst. Staining media was removed and replaced with DPBS. Compared to the vehicle-treated control, staurosporine-treated cells show a decrease in mitochondrial staining with TMRE, as well as pyknotic nuclei characteristic of apoptotic cells. Cells were imaged on a BD Pathway 435 Cell Analyzer and merged using BD Attovision software. MitoStatus TMRE has been tested on mouse data not shown . Panel 1. Flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO solid line histograms , 5 muM camptothecin Left Plot, dashed line histogram for 4 hr or 50 muM FCCP 20 min, Right Plot, dashed line histogram , and then stained with 100 nM BD Pharmingen MitoStatus TMRE 15 min, Cat. No. 564696 . Panel 2. Two-color flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO Left Plot or 5 muM camptothecin Right Plot for 4 hr, then stained with 100 nM MitoStatus TMRE 15 min . Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with APC Annexin V 15 min, Cat. No. 556419 . Co-staining shows two main populations: healthy cells that are MitoStatus TMRE-positive and Annexin V-negative, and apoptotic or dead cells that are MitoStatus TMRE-negative and APC Annexin V-positive. Compared to the DMSO-vehicle treated control, camptothecin treatment increased the MitoStatus TMRE-negative and APC Annexin V-positive population, indicating that more cells are undergoing apoptosis. Loss of mitochondrial membrane potential is an early apoptotic marker so a small population of cells in transition is MitoStatus TMRE-negative and APC Annexin V-negative. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 3. Immunofluorescent imaging of TMRE in HeLa Cells. HeLa cells were treated with 0.02 % DMSO Left Image or 1 uM staurosporine Right Image for 3 hr. Cells were stained 30 min with 200 nM MitoStatus TMRE and 5 ug/mL Hoechst. Staining media was removed and replaced with DPBS. Compared to the vehicle-treated control, staurosporine-treated cells show a decrease in mitochondrial staining with TMRE, as well as pyknotic nuclei characteristic of apoptotic cells. Cells were imaged on a BD Pathway 435 Cell Analyzer and merged using BD Attovision software. MitoStatus TMRE has been tested on mouse data not shown . Show More Panel 1. Flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO solid line histograms , 5 muM camptothecin Left Plot, dashed line histogram for 4 hr or 50 muM FCCP 20 min, Right Plot, dashed line histogram , and then stained with 100 nM BD Pharmingen MitoStatus TMRE 15 min, Cat. No. 564696 . Panel 2. Two-color flow cytometric analysis of TMRE staining in Jurkat Cells. Jurkat cells were treated with 0.025 % DMSO Left Plot or 5 muM camptothecin Right Plot for 4 hr, then stained with 100 nM MitoStatus TMRE 15 min . Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with APC Annexin V 15 min, Cat. No. 556419 . Co-staining shows two main populations: healthy cells that are MitoStatus TMRE-positive and Annexin V-negative, and apoptotic or dead cells that are MitoStatus TMRE-negative and APC Annexin V-positive. Compared to the DMSO-vehicle treated control, camptothecin treatment increased the MitoStatus TMRE-negative and APC Annexin V-positive population, indicating that more cells are undergoing apoptosis. Loss of mitochondrial membrane potential is an early apoptotic marker so a small population of cells in transition is MitoStatus TMRE-negative and APC Annexin V-negative. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 3. Immunofluorescent imaging of TMRE in HeLa Cells. HeLa cells were treated with 0.02 % DMSO Left Image or 1 uM staurosporine Right Image for 3 hr. Cells were stained 30 min with 200 nM MitoStatus TMRE and 5 ug/mL Hoechst. Staining media was removed and replaced with DPBS. Compared to the vehicle-treated control, staurosporine-treated cells show a decrease in mitochondrial staining with TMRE, as well as pyknotic nuclei characteristic of apoptotic cells. Cells were imaged on a BD Pathway 435 Cell Analyzer and merged using BD Attovision software. MitoStatus TMRE has been tested on mouse data not shown .