Experiment Overview
| Repository ID: | FR-FCM-ZZUU | Experiment name: | Figure.S2.Jurkat | MIFlowCyt score: | 95.75% |
| Primary researcher: | John Nash | PI/manager: | John Nash | Uploaded by: | John Nash |
| Experiment dates: | 2015-01-29 - 2015-01-29 | Dataset uploaded: | Oct 2016 | Last updated: | Jan 2017 |
| Keywords: | [human PBMCs] [PMA] [ionomycin] [NFAT] | Manuscripts: | [28094900] |
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| Organizations: |
Stanford University, Stanford, CA (USA)
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| Purpose: | To extend the capabilities of SLA to include transcription factors other than NFkB. In this instance, we use NFAT. | ||||
| Conclusion: | PMA/Ionomycin stimulation leads to detectable nuclear import of NFAT in Jurkat cells. | ||||
| Comments: | None | ||||
| Funding: | Cancer Biology Training Grant 5T32CA9302-35 | ||||
| Quality control: | All antibodies used in this experiment were rigorously titrated. CD45:Histone H3 interaction was our negative control, and served as baseline for background signal. dsDNA:Histone H3 was our positive control, serving as the upper limit for detection. We used both of these controls to benchmark the NFAT:dsDNA interaction. | ||||
Experiment variables
| Conditions | |
|---|---|
| · Untreated | Specimen_001_A2_A02_Untreated_1:200 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_A3_A03_Untreated_1:200 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_A4_A04_Untreated_1:200 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_A6_A06_Untreated_1:500 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_A7_A07_Untreated_1:500 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_A8_A08_Untreated_1:500 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_B2_B02_Untreated_1:1000 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B3_B03_Untreated_1:1000 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B4_B04_Untreated_1:1000 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_B6_B06_Untreated_1:2000 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B7_B07_Untreated_1:2000 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B8_B08_Untreated_1:2000 probe_NFAT:dsDNA_Untreated.fcs |
| · PMA:Ionomycin | Specimen_001_A2_A02_PMA:Ionomycin 60min_1:200 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A3_A03_PMA:Ionomycin 60min_1:200 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A4_A04_PMA:Ionomycin 60min_1:200 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_A6_A06_PMA:Ionomycin 60min_1:500 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A7_A07_PMA:Ionomycin 60min_1:500 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A8_A08_PMA:Ionomycin 60min_1:500 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B2_B02_PMA:Ionomycin 60min_1:1000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B3_B03_PMA:Ionomycin 60min_1:1000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B4_B04_PMA:Ionomycin 60min_1:1000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B6_B06_PMA:Ionomycin 60min_1:2000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B7_B07_PMA:Ionomycin 60min_1:2000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B8_B08_PMA:Ionomycin 60min_1:2000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs |
| Doses | |
|---|---|
| · 1:200 | Specimen_001_A2_A02_PMA:Ionomycin 60min_1:200 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A2_A02_Untreated_1:200 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_A3_A03_PMA:Ionomycin 60min_1:200 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A3_A03_Untreated_1:200 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_A4_A04_PMA:Ionomycin 60min_1:200 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_A4_A04_Untreated_1:200 probe_NFAT:dsDNA_Untreated.fcs |
| · 1:500 | Specimen_001_A6_A06_PMA:Ionomycin 60min_1:500 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A6_A06_Untreated_1:500 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_A7_A07_PMA:Ionomycin 60min_1:500 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A7_A07_Untreated_1:500 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_A8_A08_PMA:Ionomycin 60min_1:500 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_A8_A08_Untreated_1:500 probe_NFAT:dsDNA_Untreated.fcs |
| · 1:1000 | Specimen_001_B2_B02_PMA:Ionomycin 60min_1:1000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B2_B02_Untreated_1:1000 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B3_B03_PMA:Ionomycin 60min_1:1000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B3_B03_Untreated_1:1000 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B4_B04_PMA:Ionomycin 60min_1:1000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B4_B04_Untreated_1:1000 probe_NFAT:dsDNA_Untreated.fcs |
| · 1:2000 | Specimen_001_B6_B06_PMA:Ionomycin 60min_1:2000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B6_B06_Untreated_1:2000 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B7_B07_PMA:Ionomycin 60min_1:2000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B7_B07_Untreated_1:2000 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B8_B08_PMA:Ionomycin 60min_1:2000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B8_B08_Untreated_1:2000 probe_NFAT:dsDNA_Untreated.fcs |
| Individuals | |
|---|---|
| · CD45:Histone H3 | Specimen_001_A2_A02_PMA:Ionomycin 60min_1:200 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A2_A02_Untreated_1:200 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_A6_A06_PMA:Ionomycin 60min_1:500 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A6_A06_Untreated_1:500 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B2_B02_PMA:Ionomycin 60min_1:1000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B2_B02_Untreated_1:1000 probe_CD45:Histone H3_Untreated.fcs · Specimen_001_B6_B06_PMA:Ionomycin 60min_1:2000 probe_CD45:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B6_B06_Untreated_1:2000 probe_CD45:Histone H3_Untreated.fcs |
| · dsDNA:Histone H3 | Specimen_001_A3_A03_PMA:Ionomycin 60min_1:200 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A3_A03_Untreated_1:200 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_A7_A07_PMA:Ionomycin 60min_1:500 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_A7_A07_Untreated_1:500 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B3_B03_PMA:Ionomycin 60min_1:1000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B3_B03_Untreated_1:1000 probe_dsDNA:Histone H3_Untreated.fcs · Specimen_001_B7_B07_PMA:Ionomycin 60min_1:2000 probe_dsDNA:Histone H3_PMA:Ionomycin 60min.fcs · Specimen_001_B7_B07_Untreated_1:2000 probe_dsDNA:Histone H3_Untreated.fcs |
| · NFAT:dsDNA | Specimen_001_A4_A04_PMA:Ionomycin 60min_1:200 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_A4_A04_Untreated_1:200 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_A8_A08_PMA:Ionomycin 60min_1:500 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_A8_A08_Untreated_1:500 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_B4_B04_PMA:Ionomycin 60min_1:1000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B4_B04_Untreated_1:1000 probe_NFAT:dsDNA_Untreated.fcs · Specimen_001_B8_B08_PMA:Ionomycin 60min_1:2000 probe_NFAT:dsDNA_PMA:Ionomycin 60min.fcs · Specimen_001_B8_B08_Untreated_1:2000 probe_NFAT:dsDNA_Untreated.fcs |
