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Lsr18 machine

Manufactured by FlowJo
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The LSR18 is a flow cytometer instrument manufactured by FlowJo. It is designed to analyze and sort cells based on their physical and fluorescent properties. The LSR18 can detect up to 18 different fluorescent signals simultaneously, allowing for the comprehensive analysis of complex samples.

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Lab products found in correlation

Annexin 5 fitc and pi kit, by Thermo Fisher Scientific (3 mentions) Propidium iodide, by Merck Group (2 mentions) Pcba 1 scei plasmid, by Addgene (1 mentions) Pi solution, by Merck Group (1 mentions)

6 protocols using lsr18 machine

1

Annexin V-FITC and TMRE Assay

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Phosphatidylserine externalization was detected using an Annexin V FITC and PI kit (Thermo Fisher, Cat. No. V13242) following the manufacturer’s instructions. Briefly, cells were washed with cold PBS and resuspended in Annexin V binding buffer and stained with Annexin V and PI at room temperature and then analyzed immediately. To measure change in mitochondria membrane potential cells were treated with 200nM of tetramethylrhodamine, ethyl ester (TMRE; Abcam, Cat. No. ab113852) for 15min at 37 °C. Annexin V and TMRE-positive cells were detected using the Becton-Dickinson LSR18 machine and analysed with FlowJo version 7 software module.
Bitler B.G., Wu S., Park P.H., Hai Y., Aird K.M., Wang Y., Zhai Y., Kossenkov A.V., Vara-Ailor A., Rauscher FJ I.I.I., Zou W., Speicher D.W., Huntsman D.G., Conejo-Garcia J.R., Cho K.R., Christianson D.W, & Zhang R. (2017). ARID1A-mutated ovarian cancers depend on HDAC6 activity. Nature cell biology, 19(8), 962-973.
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2

Annexin V FITC and PI Cytometry Assay

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The Annexin V FITC and PI kit (Thermo Fisher, Cat. No. V13242) was used following the manufacturer’s instructions. Briefly, cells were washed by PBS, trypsinized, and suspended in annexin V binding buffer. Cells were stained with annexin V and PI for 15 min and then analyzed. Analysis was performed using FlowJo version 7 software module.
For cell cycle analysis, the cells were fixed, treated with bovine RNase, and then stained with propidium iodide (Sigma-Aldrich) for 15 minutes at room temperature. Samples were subjected to analysis using the Becton-Dickinson LSR18 machine and FlowJo version 7 software module.
Fukumoto T., Zhu H., Nacarelli T., Karakashev S., Fatkhutdinov N., Wu S., Liu P., Kossenkov A.V., Showe L.C., Jean S., Zhang L, & Zhang R. (2019). N6-methylation of adenosine (m6A) of FZD10 mRNA contributes to PARP inhibitor resistance. Cancer research, 79(11), 2812-2820.
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3

Annexin V-FITC and TMRE Assay

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Phosphatidylserine externalization was detected using an Annexin V FITC and PI kit (Thermo Fisher, Cat. No. V13242) following the manufacturer’s instructions. Briefly, cells were washed with cold PBS and resuspended in Annexin V binding buffer and stained with Annexin V and PI at room temperature and then analyzed immediately. To measure change in mitochondria membrane potential cells were treated with 200nM of tetramethylrhodamine, ethyl ester (TMRE; Abcam, Cat. No. ab113852) for 15min at 37 °C. Annexin V and TMRE-positive cells were detected using the Becton-Dickinson LSR18 machine and analysed with FlowJo version 7 software module.
Bitler B.G., Wu S., Park P.H., Hai Y., Aird K.M., Wang Y., Zhai Y., Kossenkov A.V., Vara-Ailor A., Rauscher FJ I.I.I., Zou W., Speicher D.W., Huntsman D.G., Conejo-Garcia J.R., Cho K.R., Christianson D.W, & Zhang R. (2017). ARID1A-mutated ovarian cancers depend on HDAC6 activity. Nature cell biology, 19(8), 962-973.
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4

Dual HR and NHEJ Reporter Assay

Cited 1 time
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The dual HR and NHEJ reporter assay was performed as previously described (Arnoult et al., 2017 (link)). Briefly, indicated cells were plated in a 6-well plates at a 50% confluence one day before transfection. Cells were transfected with 500ng of pCBA I-SceI plasmid (Addgene), 500ng of pLCN-DRR plasmid (Addgene) plasmid, and 500ng of pCAGGS-DRR-mCherry-Donor-EF1a-BFP plasmid (Addgene). After 72 hr incubation cells were collected and BFP, mCherry, and eGFP expression was detected by using the Becton-Dickinson LSR18 machine and analyzed with FlowJo version 7 software module.
Karakashev S., Fukumoto T., Zhao B., Lin J., Wu S., Fatkhutdinov N., Park P.H., Semenova G., Jean S., Cadungog M.G., Borowsky M.E., Kossenkov A.V., Liu Q, & Zhang R. (2020). EZH2 inhibition sensitizes CARM1-high, homologous recombination proficient ovarian cancers to PARP inhibition. Cancer cell, 37(2), 157-167.e6.
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5

Annexin V Apoptosis Assay

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Phosphatidylserine externalization was detected using an Annexin V FITC and PI kit following the manufacturer’s instructions. Briefly, cells were washed with cold PBS and resuspended in Annexin V binding buffer and stained with Annexin V and PI at room temperature and then analyzed immediately. Annexin V-positive cells were detected using the Becton-Dickinson LSR18 machine and analyzed with FlowJo version 7 software module.
Karakashev S., Fukumoto T., Zhao B., Lin J., Wu S., Fatkhutdinov N., Park P.H., Semenova G., Jean S., Cadungog M.G., Borowsky M.E., Kossenkov A.V., Liu Q, & Zhang R. (2020). EZH2 inhibition sensitizes CARM1-high, homologous recombination proficient ovarian cancers to PARP inhibition. Cancer cell, 37(2), 157-167.e6.
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6

Cell Cycle Analysis by Flow Cytometry

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Cells were fixed with 70% ethanol, treated with bovine RNAase for 30 min, and labeled with propidium iodide (PI) solution (Sigma). Cell suspension were incubated for 15 min at room temperature and PI staining was detected by using the Becton-Dickinson LSR18 machine, and analyzed with FlowJo version 7 software module.
Karakashev S., Fukumoto T., Zhao B., Lin J., Wu S., Fatkhutdinov N., Park P.H., Semenova G., Jean S., Cadungog M.G., Borowsky M.E., Kossenkov A.V., Liu Q, & Zhang R. (2020). EZH2 inhibition sensitizes CARM1-high, homologous recombination proficient ovarian cancers to PARP inhibition. Cancer cell, 37(2), 157-167.e6.
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