Figure 8
Immunofluorescence microscopy to examine phospho-Cav1 and β-actin localization during C. neoformans infection. The HBMEC monolayers were incubated with 106 C. neoformans strain B-4500FO2 for 0 min, 5 min and 60 min. In parallel, untreated HBMEC were used as a negative control, indicated as 0 min in the figure. β-Actin was stained with phalloidin-rhodamine conjugate (red) (second column) and the phosphor-Cav1 was stained with anti-phospho-Cav1 (Tyr-14) antibody and second antibody-FITC conjugate (green). Nuclei were stained with DAPI (blue). Bar: 15 μm. A similar study was performed in the bottom panel, in which tubulin was stained with anti-tubulin antibodies and rhodamine conjugate (second column) at the 60 min incubation time point.