Fig. 3: Selected RNA cargo mediates CXCL10 translation inhibition via the RIG-I receptor.
From: Malaria parasites both repress host CXCL10 and use it as a cue for growth acceleration
A THP-1 cells were transfected with RNA purified from: Pf ring stage, trophozoite stage, Pf-ring-stage-derived EVs or EVs derived from Pf-trophozoite-(TR) stage. In addition, all the cells were transfected with Pf gDNA. An ELISA was performed on the cell media for CXCL10 16 h post transfection. n = 3 biologically independent experiments, SEM, one-way ANOVA followed by Dunnett’s test (Pf-DNA + Ring EVs RNA - Pf-DNA ***P < 0.001). NT not treated. B THP-1 cells were transfected with Pf gDNA, NT, or RNA purified from: Pf ring stage, trophozoite stage, Pf-ring-stage-derived EVs, or Pf-trophozoite-stage-derived EVs. An ELISA was performed on the cell media for CXCL10 16 h post transfection. n = 3 biologically independent experiments, SEM, one-way ANOVA followed by Dunnett’s test, (Pf-DNA - NT ***P < 0.001, Ring-cell RNA - NT *P = 0.0405). C THP-1 cells were transfected with RNA purified from: Pf-derived EVs, Pf-derived-EVs pretreated with RNase H and Pf-derived EVs pretreated with RNase I. In addition, the cells were transfected with Pf gDNA or NT. An ELISA was performed on the cell media for CXCL10 16 h post transfection. n = 3 biologically independent experiments, SEM, one-way ANOVA followed by Dunnett’s test, (NT- Pf-DNA ***P < 0.001, Pf-DNA + Ring EVs RNA - Pf-DNA ***P < 0.001, Pf-DNA + RNase H - Pf-DNA *P = 0.0439, Pf-DNA + RNase I - Pf-DNA *P = 0.0112, Ring EVs-RNA - Pf-DNA ***P < 0.001, RNase H - Pf-DNA ***P < 0.001, RNase I - Pf-DNA ***P < 0.001). D THP-1 RIG-I KO cells were incubated with EVs harvested from ring-stage Pf (iRBCs) and then transfected with poly(dA:dT) or NT for 24 h. An ELISA assay for detecting CXCL10 was performed on cell lysates. n = 3 biologically independent experiments, SEM, one-way ANOVA followed by Dunnett’s test, (THP-1 WT- Pf EVs-dAdT ***P = 0.0000428). E Western Blot assay of CXCL10 and HSP90 (loading control) were performed on THP-1 RIG-I KO cells. The THP-1 cells were incubated with Pf-ring-stage-derived EVs, transfected with poly(dA:dT) or not treated (NT) 16 h before harvesting. Results are representative of at least three independent biological replicates. Source data are provided as a Source Data file.
