Fig. 2: FRM1 is needed for invasion while FRM2 is required for division.

a Immunofluorescence microscopy of control and rapamycin (Rap) treated FRM1-HA parasites labelled with anti-HA (green) and anti-GAP45 (red). Merge, DAPI (blue) and DIC images are also shown. Scale bars = 5 μm. b Asexual growth assay of FRM1-HA parasites following treatment with rapamycin (Rap) or DMSO (Control). The mean values and standard deviation from four experiments are plotted. An unpaired t test was performed, ***P < 0.001. c–f Rapamycin-mediated deletion of FRM1 blocks merozoite invasion of human red blood cells (RBCs). c Merozoites from FRM1-HA schizonts mock-treated with DMSO appear to invade RBCs with similar kinetics, with respect to internalisation and time to echinocytosis (EC), as other parasite lines; but were found to have a longer pre-invasion phase. d Following treatment with 100 nM Rap from ring stage, the merozoites liberated from egressing schizonts do not reinvade. e Merozoites released from Rap-treated schizonts make similar numbers of contacts (>1 s) with neighbouring RBCs as mock-treated merozoites. f Rap-treated merozoites invade significantly less efficiently than DMSO-treated merozoites. Individual data points from each cell analysed is plotted. Eight and 11 egress events were scored for the Control and Rapamycin-treated groups, respectively. The mean and standard deviation are shown. A Mann–Whitney test was performed, *P < 0.05, **P < 0.01. g Live-cell images of asexual stage transfectants. FRM2-GFP (green) puncta can be seen adjacent to the nuclei in control-treated cells. Following treatment with Rapalog (Rapa), puncta are located at the parasite membrane. Scale bar = 5 μm. h Long-term growth assay. Parasites were treated from ring stage in cycle 1 and parasitemias measured at the ring stage after 1 and 2 cycles. The data are plotted as the accumulative parasitemia over the three cycles of growth, following Rapa or Control treatment. The mean and standard error of the mean are shown, from three experiments. Unpaired t tests were performed at each cycle. P < 0.0001 for both cycle 2 and 3 Control and Rapa comparisons. Source data for this figure is provided in the Source Data files.