Figure 7: Expression of gABCG2 under the control of the CRT promoter in ΔgABCG2 parasites does not reverse gametocyte overproduction. | Nature Communications

Figure 7: Expression of gABCG2 under the control of the CRT promoter in ΔgABCG2 parasites does not reverse gametocyte overproduction.

From: A female gametocyte-specific ABC transporter plays a role in lipid metabolism in the malaria parasite

Figure 7

Live-cell fluorescence microscopy of pGLUX-6/crt-ΔPfgABCG2 transfectant was performed at asexual (a) and gametocyte (b) stages. BF, bright field. Scale bar, 5 μm. (c) The numbers of RBCs infected with 3D7-wild-type, -knockout (ΔgABCG2-07), overexpression (pGLUX-1/crt-wild type) and CRT complementation (pGLUX-6/crt-ΔPfgABCG2-07) parasites at different stages were counted by flow cytometer. At least 1,000 infected RBCs per experiment were counted. Transgenic parasite numbers relative to the wild-type numbers are shown. III–V, gametocytes stage 3 to stage 5. Gm, gametes. Means (±s.d.) are shown (n=3). (d) The expression of gABCG2 was determined by western blot performed on the synchronous GFP-tagged gABCG2 (pGREP-1-wild type), GFP-tagged endogenous complement (pGLUX-6/endo-ΔPfgABCG2) and GFP-tagged CRT complement (pGLUX-6/crt-ΔPfgABCG2) parasite samples using anti-GFP antibody. The membranes were also probed with anti-PfHSP70 antibody as a loading control. Antibody against Pfs16 was used as a marker of gametocytes. For full-sized western blots, see Supplementary Fig. 8.

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