Figure 7A

A. MRC5 cells were plated on coverslips and infected with TB40/E-mCh), UL88-STOP-mCh, or UL88-Rev-mCh at MOI of 0.05. Cells were fixed day 7 and day 9 and subjected to IF analysis: DAPI (blue) and NF-KB (green), infection (red). NF-KB translocation into the nucleus was detected via staining, and cells with nuclear NF-KB signal were classified as “NF-KB positive nuclei.” Percent of NF-KB positive nuclei over total cells counted were calculated from at least 10 fields containing 100 cells each for each sample as shown in the graph. B. MRC5 cells expressing either GFP-vector or GFP-UL88 were plated on coverslips and infected with mCherry tagged WT or UL88-STOP TB40/E virus at MOI of 0.05. Cells were fixed day 7 and day 9 and subjected to IF analysis: DAPI (blue), GFP (green) and NF-KB (grey), infection (red). NF-KB translocation into the nucleus was detected via staining, and cells with nuclear NF-KB signal were classified as “NF-KB positive nuclei.” Percent of NF-KB positive nuclei over total cells counted were calculated from at least 10 fields containing 100 cells each for each sample as shown in the graph.A. 将MRC5细胞铺于盖玻片上，以感染复数（Multiplicity of Infection, MOI）为0.05的TB40/E-mCh、UL88-STOP-mCh或UL88-Rev-mCh病毒分别感染细胞。分别于感染后第7天和第9天固定细胞，进行免疫荧光（Immunofluorescence, IF）分析：采用4',6-二脒基-2-苯基吲哚（4',6-Diamidino-2-Phenylindole, DAPI，蓝色）、核因子κB（Nuclear Factor-kappa B, NF-κB，绿色）以及感染标记（红色）进行染色。通过染色检测NF-κB向细胞核内的转位情况，将带有细胞核NF-κB信号的细胞归类为“NF-κB阳性细胞核”。计算每个样本中NF-κB阳性细胞核占总计数细胞的百分比，每个样本至少统计10个视野，每个视野计数不少于100个细胞，结果如图所示。B. 将表达绿色荧光蛋白（Green Fluorescent Protein, GFP）空载载体或GFP-UL88的MRC5细胞铺于盖玻片上，以感染复数（MOI）为0.05的mCherry标记野生型（Wild Type, WT）或UL88-STOP TB40/E病毒感染细胞。分别于感染后第7天和第9天固定细胞，进行免疫荧光（IF）分析：采用DAPI（蓝色）、GFP（绿色）、NF-κB（灰色）以及感染标记（红色）进行染色。通过染色检测NF-κB向细胞核内的转位情况，将带有细胞核NF-κB信号的细胞归类为“NF-κB阳性细胞核”。计算每个样本中NF-κB阳性细胞核占总计数细胞的百分比，每个样本至少统计10个视野，每个视野计数不少于100个细胞，结果如图所示。