Fig. 5.
Depletion of GAPM1a causes rapid changes in vacuolar organisation, cell morphology, and 3D motility. a Live video microscopy of intracellular parasites co-expressing GAPM1a-AID and mTagRFP-T_TubA1. Acute depletion of GAPM1a-AID led to the loss of the characteristic rosette organisation. Graph represents mean ± SD for n = 16 vacuoles per condition; **p < 0.005, ****p < 0.00005, Student’s t test with Holm–Sidak correction. b Length of individual, non-dividing parasites was measured every 14 min. Graph represents mean ± SD for n = 21 parasites per condition, * p < 0.05, ***p < 0.0005, Student’s t test with Holm–Sidak correction. Representative images shown for (a, b). Scale bars are 5 μm. c Comparison of parasite length following extracellular GAPM1a depletion or microtubule depolymerisation after incubating TKO parasites for 4 h at 4° C. Box plots for n = 561 (NT, 37 °C), 348 (NT, 4°C), 326 (4 h), 616 (TKO, 37 °C), 266 (TKO 4 °C) parasites per condition aggregated from three independent experiments. d Parasite volume estimated from over 8000 parasites per sample by Coulter counter. Asterisk indicates treatments performed before mechanical release of parasites from host cells. Box plots for n = 6 (37 °C, 4 h*), 7 (37 °C, NT*), 8 (37 °C, NT and 37 °C,TKO), 7 (4 °C, TKO) biological replicates; p values from two-tailed Student’s t test. e Buoyant mass of parasites was quantified using SMR. Box plots for n = 479 (NT), 509 (4 h), 442 (TKO 37oC), 455 (TKO 4oC) aggregated from three independent experiments; p values from two-tailed Student’s t test. f Maximum intensity projections of GAPM1a-AID parasite 3D motility either untreated or pre-treated for 4 h with IAA. Scale bar is 100 μm. g Mean displacement measured for at least 150 parasite per experiment. Graph shows mean ± SD for n = 5 biological replicates; p values from two-tailed Student’s t test. All box plots represent median and 25th and 75th percentiles and whiskers are at 10th and 90th percentiles. Source data are provided as a Source Data file