Fig 1: Arp2/3 is required for endocytic trafficking of transferrin in C. burnetii infected cells.(A-C) Two dpi Vero cells treated with CK-666 or DMSO for 24 hr were incubated with 488-transferrin (488Tfr) for 1 hr, washed with PBS, then fixed and immunostained for CD63 and C. burnetii. CK-666 inhibits trafficking of 488Tfr to CCVs, decreasing the amount of 488Tfr on the CCV and colocalization with CCV CD63 clusters. Graphs represent the means ± SD of = 60 cells from 3 independent experiments. Statistical significance was determined by the Student’s t-test (****P <0.0001). NMII, C. burnetii Nine Mile phase II strain. Scale bar, 5 µm.
Fig 2: Arp2/3 is needed for CCV biogenesis.(A and B) CK-666 treatment prevents CCV expansion. Vero cells at 24 hr post-infection were treated with CK-666 or chloramphenicol for 2 days. Cells were fixed and fluorescently stained for CD63 and F-actin. (C and D) CK-666 treatment causes CCV collapse. Vero cells at 2 dpi were treated with CK-666 for 24 hr, then fixed along with the corresponding 3 dpi DMSO-treated control or washed to remove CK-666 and allowed an additional 24 hr recovery before fixation. Cells were fluorescently stained for F-actin and CD63. Histograms depict the means of the percentage of cells with normal CCV expansion or CCV area ± SD of = 60 cells for at least 3 independent experiments. Statistical significance was determined by the Student’s t-test (****P <0.0001). NMII, C. burnetii Nine Mile phase II strain. Scale bar, 5 µm.
Supplier Page from Abcam for Anti-Arp2 (phospho T237 + T238) antibody