Disruption of actin regulators reduces the entry and trafficking of KSHV particles to the perinuclear region.

(A and E) HUVEC were treated with chemicals to inhibit Rho GTPase function (A) or Arp2/3 complex activity (E) at the indicated doses for 1 h prior to inoculation with KSHV. Cells were inoculated with KSHV in the presence of inhibitors, fixed at 4 hpi, and stained for Orf65+ viral particles (red) and nuclei (blue). (B and F) The total number of nuclei bearing at least one Orf65+ particle was determined. In parallel experiments, cells were subjected to the same treatments and evaluated for viability by PI staining. (C and G) The total number of Orf65+ particles docked at each nucleus was determined. (D and H) Disruption of the actin cytoskeleton by CdTB (D) and wiskostatin (H). HUVEC were treated with the inhibitors as described in A and E, and stained for the actin cytoskeleton with AlexaFluor 488-phalloidin.

(A与E组) 人脐静脉内皮细胞（Human Umbilical Vein Endothelial Cells，HUVEC）先以指定浓度的化学试剂预处理1小时，以抑制Rho鸟苷三磷酸酶（Rho GTPase）功能(A)或Arp2/3复合物（Arp2/3 complex）活性(E)，随后接种卡波西肉瘤相关疱疹病毒（Kaposi's Sarcoma-Associated Herpesvirus，KSHV）。实验全程保留抑制剂，于感染后4小时（hours post infection，hpi）固定细胞，随后对Orf65阳性病毒颗粒（红色荧光标记）与细胞核（蓝色荧光标记）进行染色。
(B与F组) 统计至少携带1个Orf65阳性颗粒的细胞核总数。平行实验中，细胞接受相同处理后，通过碘化丙啶（Propidium Iodide，PI）染色评估细胞活力。
(C与G组) 统计每个细胞核表面锚定的Orf65阳性颗粒总数。
(D与H组) 分别展示CdTB与Wiskostatin对肌动蛋白细胞骨架的破坏效果。将HUVEC按照(A)与(E)所述方法用抑制剂处理后，采用AlexaFluor 488-鬼笔环肽（AlexaFluor 488-phalloidin）对肌动蛋白细胞骨架进行染色。