A quorum sensing-independent path to stumpy development in Trypanosoma brucei
Fig 4
Silencing of the ES-resident VSG is independent of ES-attenuation.
The transcriptional status of the active ES of growth arrested (left) and proliferating (right) ectopic VSG 121 overexpressors was monitored. (A, B) The GFPESpro reporter was used to detect transcripts from the ES promotor region (GFP) and (C, D) ESAG6 was measured as an example for a native ES transcript. All data points reflect measurements on the single cell level using mRNA FISH (Affymetrix). The signal intensity in deconvolved, summed slice projections (100 images, z-step 100 nm) was measured with Image J and is represented as relative fluorescent unit (RFU). Two different induction times (24 and 48 h) were analyzed, and non-induced long slender (0 h) or density-induced short stumpy cells (st) served as controls. Only parasites in G1-phase of the cell cycle were analyzed. The magenta bars are means ± SD (n > 100). Statistical analysis was conducted using an unpaired t-test (not significant (n.s.) p-value > 0.05; ** p-value < 0.01; *** p-value < 0.001).