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Experiment Overview

Repository ID: FR-FCM-Z5R4 Experiment name: Tse5-CT MIFlowCyt score: 96.00%
Primary researcher: Jon Altuna PI/manager: Jon Altuna Uploaded by: Jon Altuna
Experiment dates: 2021-03-22 - 2022-03-24 Dataset uploaded: Oct 2022 Last updated: Oct 2022
Keywords: [Pseudomonas putida] [Pseudomonas aeruginosa] [T6SS] [Tse5] [toxin] Manuscripts:
Organizations: Instituto Biofisika, Leioa, (España)
Purpose: Evaluating the effefct of Tse5-CT on Pseudomonas putida EM383 cells.
Conclusion: Tse5-CT causes cell membrane depolarization when expressed in Pseudomona putida EM383 cells. Tse5-CT effect can be neutralized when Tsi5 inmunity protein is expressed in P. putida EM383 strain.
Comments: None
Funding: Not disclosed
Quality control: Positive permeabilitation (SYTOX™ Red) control, Positive depolarization (DiBAC4(3) (Bis-(1,3-Dibutylbarbituric Acid)Trimethine Oxonol)) control and negative unstained cell control


Experiment variables

Sample Type
· Positive Permeabilitation control Heat Shock (1).fcs · Heat Shock (2).fcs · Heat Shock (3).fcs
· Positive Depolarization Control Polymixyn B (1).fcs · Polymixyn B (2).fcs · Polymixyn B (3).fcs
· Cells gating Cell gating.fcs
· Tse5-CT vs sp-Tse5-CT Tse5-CT (1).fcs · Tse5-CT (2).fcs · Tse5-CT (3).fcs · sp-Tse5-CT (1).fcs · sp-Tse5-CT (2).fcs · sp-Tse5-CT (3).fcs
· Neutralization Tse5-CT & Tsi5 (+) (1).fcs · Tse5-CT & Tsi5 (+) (2).fcs · Tse5-CT & Tsi5 (+) (3).fcs
· Negative Control Empty Vector (1).fcs · Empty Vector (2).fcs · Empty Vector (3).fcs · Tse5-CT + Tsi5 (-) (1).fcs · Tse5-CT + Tsi5 (-) (2).fcs · Tse5-CT + Tsi5 (-) (3).fcs

Conditions
· No induction Cell gating.fcs · Empty Vector (1).fcs · Empty Vector (2).fcs · Empty Vector (3).fcs · Tse5-CT + Tsi5 (-) (1).fcs · Tse5-CT + Tsi5 (-) (2).fcs · Tse5-CT + Tsi5 (-) (3).fcs
· 1 mM m-toluenic acid induction Tse5-CT (1).fcs · Tse5-CT (2).fcs · Tse5-CT (3).fcs · sp-Tse5-CT (1).fcs · sp-Tse5-CT (2).fcs · sp-Tse5-CT (3).fcs
· 1 mM m-toluenic and 100 nM IPTG co-induction Tse5-CT & Tsi5 (+) (1).fcs · Tse5-CT & Tsi5 (+) (2).fcs · Tse5-CT & Tsi5 (+) (3).fcs
· Heat Shock Heat Shock (1).fcs · Heat Shock (2).fcs · Heat Shock (3).fcs
· Antibiotic depolarization Polymixyn B (1).fcs · Polymixyn B (2).fcs · Polymixyn B (3).fcs

Doses
· None Cell gating.fcs · Empty Vector (1).fcs · Empty Vector (2).fcs · Empty Vector (3).fcs · Heat Shock (1).fcs · Heat Shock (2).fcs · Heat Shock (3).fcs · Polymixyn B (1).fcs · Polymixyn B (2).fcs · Polymixyn B (3).fcs · Tse5-CT & Tsi5 (+) (1).fcs · Tse5-CT & Tsi5 (+) (2).fcs · Tse5-CT & Tsi5 (+) (3).fcs · Tse5-CT (1).fcs · Tse5-CT (2).fcs · Tse5-CT (3).fcs · Tse5-CT + Tsi5 (-) (1).fcs · Tse5-CT + Tsi5 (-) (2).fcs · Tse5-CT + Tsi5 (-) (3).fcs · sp-Tse5-CT (1).fcs · sp-Tse5-CT (2).fcs · sp-Tse5-CT (3).fcs

Timepoints
· Same Cell gating.fcs · Empty Vector (1).fcs · Empty Vector (2).fcs · Empty Vector (3).fcs · Heat Shock (1).fcs · Heat Shock (2).fcs · Heat Shock (3).fcs · Polymixyn B (1).fcs · Polymixyn B (2).fcs · Polymixyn B (3).fcs · Tse5-CT & Tsi5 (+) (1).fcs · Tse5-CT & Tsi5 (+) (2).fcs · Tse5-CT & Tsi5 (+) (3).fcs · Tse5-CT (1).fcs · Tse5-CT (2).fcs · Tse5-CT (3).fcs · Tse5-CT + Tsi5 (-) (1).fcs · Tse5-CT + Tsi5 (-) (2).fcs · Tse5-CT + Tsi5 (-) (3).fcs · sp-Tse5-CT (1).fcs · sp-Tse5-CT (2).fcs · sp-Tse5-CT (3).fcs

Individuals
· Pseudomonas putida EM383 Cell gating.fcs · Empty Vector (1).fcs · Empty Vector (2).fcs · Empty Vector (3).fcs · Heat Shock (1).fcs · Heat Shock (2).fcs · Heat Shock (3).fcs · Polymixyn B (1).fcs · Polymixyn B (2).fcs · Polymixyn B (3).fcs · Tse5-CT & Tsi5 (+) (1).fcs · Tse5-CT & Tsi5 (+) (2).fcs · Tse5-CT & Tsi5 (+) (3).fcs · Tse5-CT (1).fcs · Tse5-CT (2).fcs · Tse5-CT (3).fcs · Tse5-CT + Tsi5 (-) (1).fcs · Tse5-CT + Tsi5 (-) (2).fcs · Tse5-CT + Tsi5 (-) (3).fcs · sp-Tse5-CT (1).fcs · sp-Tse5-CT (2).fcs · sp-Tse5-CT (3).fcs

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