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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: Prolonged activation of cytomegalovirus early gene e1-promoter exclusively in neurons during infection of the developing cerebrum

Fig. 3

at Detection of EGFP, MCMV-E1 and MCMV-M45 in rMCMV1373- or 448-infected MEF and primary neuronal cultures. Either rMCMV was allowed to infect both cell cultures at a MOI of 1. Virus-infected MEF and primary neuronal cultures were sampled at 2 and 4 dpi, respectively. ad rMCMV1373-infected MEFs. eh rMCMV448-infected MEFs. in rMCMV1373-infected primary neuronal culture. ot rMCMV448-infected primary neuronal culture. In each rMCMV-infected culture, phase contrast (a, e, i, o) and fluorescence views of EGFP (b, f, j, n, p, t), immunofluorescence for MCMV-E1 (red) (c, g, k, q) and MCMV-45 (red) (m, s) and merged images of EGFP and MCMV-E1 (d, h, l, r) are shown. Arrow heads in pt indicate EGFP+ cells. Scale bar: 50 µm. u, v Comparison of the numbers of EGFP+- or MCMV-E1+ cells in MEF (u) and primary neuronal cultures (v) infected with rMCMV1373 or rMCMV448. MEF and primary neuronal cultures were infected with rMCMVs at three different MOIs and analyzed at 2 and 4 dpi, respectively. Additionally, in primary neuronal cultures (v) the numbers of cells doubly positive for EGFP and NeuN as shown in Fig. 4 were counted. The cell counts of 3 different fields (292 × 220 µm; 0.064 mm2/a field) were averaged in each coverslip. Mean ± SEM of 3 different coverslips in each experimental group are shown (E1+, white bar; EGFP+, gray bar; EGFP+/NeuN+, black bar). Averaged numbers of total cells as DAPI+ nuclei were 339 ± 31/0.01 mm2 and 305 ± 20/0.01 mm2, in virus-infected MEF and primary neuronal cultures, respectively. The averaged percentage of NeuN+ cells in primary neuronal cultures infected with rMCMVs was 87.6 ± 3.6%. *p < 0.01 vs. corresponding rMCMV1373-infected culture in each experimental group by an unpaired 2-tailed Student’s t-test

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