Labtek permanox four well chamber slides

Mock- or KSHV-infected cells were seeded on LabTek Permanox four-well chamber slides (Fisher Scientific) and fixed with 4% (wt/vol) paraformaldehyde in phosphate-buffered saline. Immunofluorescence was performed as described previously [54 (link)]. Briefly, cells were incubated in Tris-Buffered Saline (20mM Tris, 150mM NaCl, pH 7.6; TBS) containing 1% normal goat serum followed by incubation with primary antisera at a dilution of 1:1000 (rabbit anti-LANA, a kind gift from the Ganem lab) diluted in TBS containing 1% BSA for 1 h. Cells were then incubated with fluor-conjugated secondary antibodies (Molecular Probes/Invitrogen) for 1 h. Cells were mounted in medium containing DAPI (4’,6’-diamidino-2-phenylindole) before being viewed under a Keyence BZ-X710.

Mock- or KSHV-infected cells were seeded on LabTek Permanox four-well chamber slides (Fisher Scientific) and fixed with 4% (wt/vol) paraformaldehyde in phosphate-buffered saline. Immunofluorescence was performed as described previously [27 (link)]. Briefly, cells were incubated in Tris-Buffered Saline (20 mM Tris, 150 mM NaCl, pH 7.6; TBS) containing 1% normal goat serum followed by incubation with primary antisera at a dilution of 1:1,000 (rabbit anti-LANA) diluted in TBS containing 1% BSA for 1 hour. Cells were then incubated with fluor-conjugated secondary antibodies (Molecular Probes/Invitrogen) for 1 hour. Cells were mounted in medium containing DAPI (4',6'-diamidino-2-phenylindole) before being viewed under a Nikon Eclipse E400 microscope.