Culturing Trypanosoma brucei Parasites

Procyclic PF427 parasites were cultured in SDM79 medium supplemented with 10% fetal calf serum (FCS) at 27°C. Parasites were treated with 20 μM I-BET151 (Sigma-Aldrich). Parasites were counted every 48 h and diluted to a concentration of 2 million/mL in fresh medium with fresh drug every 2 days. EP1/GFP procyclic reporter parasites were generated as described in reference 30 (link). EP1/GFP bloodstream parasites (30 (link)) and single-marker (SM) parasites (77 (link)) (used as controls for verification of the EP1/GFP reporter construct) were grown in HMI9 supplemented with 10% FCS and 10% Serum Plus at 37°C and 5.0% CO₂.

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Ashby E.C., Havens J.L., Rollosson L.M., Hardin J, & Schulz D. (2023). Chemical Inhibition of Bromodomain Proteins in Insect-Stage African Trypanosomes Perturbs Silencing of the Variant Surface Glycoprotein Repertoire and Results in Widespread Changes in the Transcriptome. Microbiology Spectrum, 11(3), e00147-23.

Publication 2023

I-BET151

Bloodstream Drug Fetal calf serum I bet151 Parasites Serum

Corresponding Organization : Harvey Mudd College

Other organizations : Pomona College

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Variable analysis

independent variables

I-BET151 (Sigma-Aldrich) at 20 μM concentration

dependent variables

Parasite count every 48 hours

control variables

Procyclic PF427 parasites were cultured in SDM79 medium supplemented with 10% fetal calf serum (FCS) at 27°C
Parasites were diluted to a concentration of 2 million/mL in fresh medium with fresh drug every 2 days

controls

Positive control: EP1/GFP procyclic reporter parasites
Negative control: EP1/GFP bloodstream parasites and single-marker (SM) parasites

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