Figure 3
Monitoring of the binding of PfApicortin with α-tubulin-I and β-tubulin by immuno-precipitation, ELISA and SPR. (A) Detection of α-tubulin-I in western blotting after pulling down from parasite lysate with recombinant apicortin (Supplementary Fig. S10A), (B) Detection of β-tubulin in western blotting after pulling down from Pf3D7 lysate using recombinant apicortin (Supplementary Fig. S10B), (C) and (D) Confirmation of the presence of recombinant apicortin bound on the bead used for pull down assay (Supplementary Fig. S10C,D), (E) Graph showing indirect ELISA data showing interaction between PfApicortin and Pfα-tubulin-I with increasing titers of recombinant Pfα-tubulin-I overlaid on apicortin coated surface (x axis indicates amount of overlaid α-tubulin-I in ng), (F) Graph showing indirect ELISA data showing interaction between PfApicortin and Pfβ-tubulin with increasing titers of recombinant Pfβ-tubulin overlaid on apicortin coated surface (x axis indicates amount of overlaid β-tubulin in ng), (G) Graph showing surface plasmon resonance data where increasing concentrations of α-tubulin-I (in µM) was injected over the surface containing immobilized apicortin, (H) Graph showing surface plasmon resonance data where increasing concentrations of β-tubulin (in µM) was injected over the surface containing immobilized apicortin (analysis of spectra using Autolab ESPRIT kinetic evaluation software (https://www.metrohm.com/en-in/products/more-products/kei/). Data are represented as mean ± SD of at least three independent experiments.
