Fig. 1
Characterization of HSV-1-infected primary cortical neurons (PCC). a Murine PCCs were infected with HSV-1(17+)LoxpCMVGFP (MOI 10) at DIV5 and compared with mock-infected control cells 16 hpi. Cells were stained against the neuronal marker βIII-tubulin (βIII-tub), the astrocytic marker glial fibrillary acidic protein (GFAP), the oligodendrocyte transcription factor (olig-2), and allograft inflammatory factor (Aif1/Iba-1) as a marker for microglia. b Cell type composition of the mock infected PCCs. c Percentage of HSV-1 positive cells 6 and 16 hpi defined for each cell type. Bars show mean ± SEM (n = 3) with a two-way ANOVA and a Holm-Sidak’s multiple comparison test (**p < 0.01, ***p < 0.001 compared to 6 hpi astrocytes, ###p < 0.001 compared to 16 hpi astrocytes). d The astrocytes in the PCCs were HSV-1(17+)LoxpCMVGFP infected (MOI 10) and analyzed 6 hpi and 16 hpi via GFAP staining. e–g GFAP positive astrocytes were characterized using the automated cell image analysis software CellProfiler. e The area of HSV-1 negative and HSV-1-positive astrocytes was measured within mock control and HSV-1-infected PCCs. f Compactness of infected and non-infected astrocytes. g Classification of HSV-1 positive and HSV-1 negative astrocytes depending on the area of the cell body related to the total astrocyte area (large > 1000 μm2, medium 1000 μm2 ≥ × ≤ 500 μm2, small < 500 μm2). Sidak’s multiple comparison tests refer to mock-infected control astrocytes of the same size-class. h–j mRNA levels of A1/A2 markers were quantified by qRT-PCR in PCCs 6 and 16 hpi. All bars show mean ± SEM (n = 3) with a two-way ANOVA (e–g) and a one-way ANOVA (h–j) followed by Sidak’s multiple comparison test (****p < 0.0001, **p < 0.01, *p < 0.05)