Cell cycle profiles of KSHV positive and negative cells treated with nocodazole showed that KSHV is capable of releasing nocodazole induced G2/M block though LANA.

<p>(A) Two KSHV positive cells (BC3 and JSC-1), and a KSHV negative cells (BJAB) were treated with or without different concentration of nocodazole (200 ng/mL, 300 ng/mL, 500 ng/mL or 1 µg/mL) for 24 hours. The cells were then harvested, stained with PI and their cell cycle profiles were determined by flow cytometry. Nocodazole treated BJAB cells showed an increase in the proportion of cells at G2/M, indicating a block, whereas BC3 and JSC-1 showed no block in G2/M phase. Results are indicated as percentages of cells in all cell cycle phases. Mean and standard deviations were derived from 3 independent experiments. (B) Western blot shows the expression of LANA in KSHV positive cells (BC-3 and JSC-1) but not in KSHV negative cells (BJAB). Histone-H3 was taken as loading control. (C) BJAB cells transfected with the pA3M vector or pA3M-LANA and were treated with nocodazole (200 ng/mL) for 24 hours. The cells were then harvested and DNA cell cycle distribution patterns were determined as above. Results in the untreated and treated cells are shown as the percentage of cells in all cell cycle phases. Mean and standard deviations were derived from 3 independent experiments. (D) Western blot shows the expression of LANA in the BJAB cells transfected with pA3M-LANA. Histone-H3 was taken as loading control. The result shown is the representative of 3 independent experiments. +, present; −, absent.