Fig. 4.

Antibodies raised against hexon fragments reveal variable cross-reactivity against hexon proteins in Western immunoblotting. Lysates from HeLa cells infected with thirteen HAdVs representing the four species B, C, D and E and CMT93 lysates with all three different MAdVs were analyzed. Uninfected HeLa or CMT93 cells were included as negative controls, respectively. The blots were stained with the HAdV-B3 hexon Ab (a), the MAdV-1 hexon Ab (b), or the MAdV-2 hexon Ab (c). In addition, all blots were immunostained using a (HAdV) cross-reactive anti-pIIIa antibody to check for efficient infection, and an actin antibody for loading control. It was noticed that the actin signal in mouse cells infected with MAdVs strongly faded in the course of infection. d A second set of CMT93 cells infected with all three MAdVs at about five-fold higher virus input were harvested 48, 72 and 96 h post infection. The blot was immunostained using the MAdV-1 and MAdV-2 hexon Ab and GAPDH loading control antibody. The same lysates were stained for protein content using PageBlue. Marker proteins of 118, 85, 48, 34, 26 and 20 kDa were included in the first lane