IF microscopy of Ad-infected 293 cells at 48 h pi.

<p>(A–D) and (H–J). Conventional IF microscopy using anti-fiber monoclonal antibody TB5 and secondary Alexa Fluor®633-labeled anti-mouse IgG. (A), Ad5WTFib; (B), Ad5PbEGD; (C), Ad5H508A; (D), Ad5(HI)RGD4C (with DAPI counterstaining). (E–G), Confocal analysis of Ad5ßGalWTFib-infected 293 cells with TB5. (E), Sample reacted with TB5 Alexa Fluor®633-labeled anti-mouse IgG; (F), sample reacted with anti-penton base antibody and secondary FITC-labeled anti-rabbit IgG; (G), image overlay with DAPI nuclear staining. (H–J), Kinetics of appearance of intranuclear crystals in Ad-infected cells. 293 cells infected with Ad5WTFib were harvested at different times pi and reacted with TB5 and Alexa Fluor®633-labeled anti-mouse IgG, with DAPI counterstaining. (H), 12 h pi; (I), 16 h pi; (J), 20 h pi. Note that IF pictures are presented at low magnification to show large field of view including several crystal-containing cells (A, B ; H–J), or at higher magnification to show the mass and number of crystals within a single nucleus (C–G).