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facs sorting  (AMS Biotechnology)


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    AMS Biotechnology facs sorting
    (A-B) Generation of HLA-A2 <t>+</t> <t>Luciferase</t> + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) <t>FACS</t> analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
    Facs Sorting, supplied by AMS Biotechnology, used in various techniques. Bioz Stars score: 98/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/facs+sorting/bio_rxiv__64898__2026__04__21__719872-204-26-20?v=AMS+Biotechnology
    Average 98 stars, based on 26 article reviews
    facs sorting - by Bioz Stars, 2026-07
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    Images

    1) Product Images from "Multifaceted immune resistance landscapes in human oligodendrocytes protect against cytotoxic T cells and are dysregulated in MS brain cell subsets"

    Article Title: Multifaceted immune resistance landscapes in human oligodendrocytes protect against cytotoxic T cells and are dysregulated in MS brain cell subsets

    Journal: bioRxiv

    doi: 10.64898/2026.04.21.719872

    (A-B) Generation of HLA-A2 + Luciferase + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) FACS analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
    Figure Legend Snippet: (A-B) Generation of HLA-A2 + Luciferase + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) FACS analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.

    Techniques Used: Luciferase, Expressing, Transfection, Plasmid Preparation, Transduction, Control, Live Cell Imaging, Quantitative RT-PCR, Cytotoxicity Assay, Co-Culture Assay, Cell Culture, Selection, Knockdown, Two Tailed Test



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    (A-B) Generation of HLA-A2 <t>+</t> <t>Luciferase</t> + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) <t>FACS</t> analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
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    Eppendorf AG facs pinsgfp entpd3 sorting pinsgfp clusters
    (A-B) Generation of HLA-A2 <t>+</t> <t>Luciferase</t> + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) <t>FACS</t> analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
    Facs Pinsgfp Entpd3 Sorting Pinsgfp Clusters, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fluorescence activated cell sorting facs buffer  (Thermo Fisher)
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    Thermo Fisher fluorescence activated cell sorting facs buffer
    (A-B) Generation of HLA-A2 <t>+</t> <t>Luciferase</t> + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) <t>FACS</t> analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
    Fluorescence Activated Cell Sorting Facs Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    facs sorting  (Sony Biotechnology)
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    (A-B) Generation of HLA-A2 <t>+</t> <t>Luciferase</t> + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) <t>FACS</t> analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.
    Facs Sorting, supplied by Sony Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fluorescence activated cell sorting facs flow cytometer  (Danaher Inc)
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    Danaher Inc fluorescence activated cell sorting facs flow cytometer
    Effects of HC on the expression of proinflammatory macrophage (CD80) and anti-inflammatory macrophage (CD163) surface markers in RAW264.7 cells. A: Percentage of CD80(−)/CD163(−) (M0) macrophages. B: Percentage of CD80(+)/CD163(−) (M1) macrophages. C: Percentage of CD80(−)/CD163(+) (M2) macrophages. D: Representative <t>FACS</t> plots of each experimental group. The data are presented as the means ± SDs (n = 3 for each treatment group). ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001; ns: not significant.
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    Image Search Results


    (A-B) Generation of HLA-A2 + Luciferase + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) FACS analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.

    Journal: bioRxiv

    Article Title: Multifaceted immune resistance landscapes in human oligodendrocytes protect against cytotoxic T cells and are dysregulated in MS brain cell subsets

    doi: 10.64898/2026.04.21.719872

    Figure Lengend Snippet: (A-B) Generation of HLA-A2 + Luciferase + human oligodendrocytic cell line MO3.13-A2-Luc. (A-B) FACS analysis of the (A) HLA-A2 expression on wildtype (WT) MO3.13 cells (left), MO3.13-A2 cells 2 weeks after transfection with HLA-A2 expression vector (middle), and MO3.13-A2 cells after the FACS sorting for HLA-A2 (right). (B) GFP expression on MO3.13-A2-Luc cells after lentiviral transduction (left), GFP expression on FACS sorted MO3.13-A2-Luc cells (middle) and HLA-A2 expression on MO3.13-A2-Luc cells (right). Grey histograms represent the isotype control and untransduced cells for the analysis of HLA-A2 and GFP expression respectively. (C-E) Optimization of siRNA reverse transfection protocol for MO3.13-A2-Luc cells. (C) Real-time live cell imaging of MO3.13-A2-Luc cells transfected either with a non-targeting siRNA control (Scr) or with a siRNA cocktail targeting genes essential for cell survival (cell death siRNA, siCD). Transfected cells were imaged via the IncuCyte SX5 system for 72 h and real-time % cell confluency was quantified. (D) RT-qPCR analysis of PDL1 mRNA expression in MO3.13-A2-Luc cells transfected either with Scr siRNA or pool of 4 non-overlapping siRNAs targeting PDL1 (siCD274). Results are presented as fold change compared to the Scr after β-actin mRNA normalization. (E) FACS analysis of PD-L1 surface expression on MO3.13-A2-Luc cells transfected either with Scr or PD-L1 specific siRNA pool. Left: representative histograms indicating % PD-L1 expression, right: mean fluorescent intensity (MFI) of PD-L1 from 2 independent FACS data. (F) Phenotypic characterization of FluTC. The expression of CD4 and CD8 was determined on CD3 + FluTC, whereas the expression of effector and memory markers CD45RO and CD62L and co-inhibitory immune checkpoint molecules PD-1, LAG-3 and TIM-3 was determined on CD3 + CD8 + FluTC. (Tn: naïve T cells, Tcm: central memory T cells, Teff: terminal effector T cells, Tem: effector memory T cells). Grey histograms represent the isotype control. (G) Luciferase-based cytotoxicity assay to optimize the co-culture conditions for MO3.13-A2-Luc and FluTC. MO3.13-A2-Luc were pulsed with serial dilutions of the flu-peptide and co-cultured with FluTC. After 20 h of co-culture, remaining relative luciferase units (RLU) were measured. RLU measured in pulsed samples were normalized to RLU of unpulsed (no peptide) control. Statistical significance was calculated compared with no peptide control. (H) Selection of cytotoxicity and viability controls for the HTP screen. MO3.13-A2-Luc cells were transfected with the indicated siRNAs. After 72 h of transfection cells were pulsed with 0,01 µg/ml flu-peptide and co-cultured either with FluTC or treated with control T cell medium (CM). 20 h following co-culture, remaining RLU was measured. The RLU of each sample was normalized to the Scr1 control in the cytotoxicity (FluTC) or viability (CM) setting to determine the impact of gene knockdown. (A-G) Representative data of at least 2 independent experiments. (H) Cumulative data of three independent experiments each performed with four replicates per sample. (D, G-H) Graphs show mean +/- SD. p-values were calculated using two-tailed student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.005, **** = p < 0.001.

    Article Snippet: MO3.13-A2-Luc cells were generated after transduction of MO3.13-A2 cells with lentiviral particles encoding firefly luciferase and GFP (GenTarget Inc – Amsbio, #LVP020 (CMV-Luciferase (firefly)-2A-GFP (Puro)) and FACS sorting.

    Techniques: Luciferase, Expressing, Transfection, Plasmid Preparation, Transduction, Control, Live Cell Imaging, Quantitative RT-PCR, Cytotoxicity Assay, Co-Culture Assay, Cell Culture, Selection, Knockdown, Two Tailed Test

    Effects of HC on the expression of proinflammatory macrophage (CD80) and anti-inflammatory macrophage (CD163) surface markers in RAW264.7 cells. A: Percentage of CD80(−)/CD163(−) (M0) macrophages. B: Percentage of CD80(+)/CD163(−) (M1) macrophages. C: Percentage of CD80(−)/CD163(+) (M2) macrophages. D: Representative FACS plots of each experimental group. The data are presented as the means ± SDs (n = 3 for each treatment group). ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001; ns: not significant.

    Journal: Journal of Traditional and Complementary Medicine

    Article Title: Anti-inflammatory and polarization-modulating effects of Houttuynia cordata in LPS-stimulated RAW264.7 macrophages

    doi: 10.1016/j.jtcme.2025.05.001

    Figure Lengend Snippet: Effects of HC on the expression of proinflammatory macrophage (CD80) and anti-inflammatory macrophage (CD163) surface markers in RAW264.7 cells. A: Percentage of CD80(−)/CD163(−) (M0) macrophages. B: Percentage of CD80(+)/CD163(−) (M1) macrophages. C: Percentage of CD80(−)/CD163(+) (M2) macrophages. D: Representative FACS plots of each experimental group. The data are presented as the means ± SDs (n = 3 for each treatment group). ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001; ns: not significant.

    Article Snippet: After the cells were fixed with 4 % paraformaldehyde, phagocytic uptake was analyzed via a Fluorescence-activated cell sorting (FACS) flow cytometer (CytoFLEX S, Backman, USA).

    Techniques: Expressing