Journal: Cells

Article Title: Subviral Dense Bodies of Human Cytomegalovirus Induce an Antiviral Type I Interferon Response.

doi: 10.3390/cells11244028

Figure Lengend Snippet: Figure 5. Western blot analysis of the IRG expression in endothelial cells (EC) after DB-exposure. ECs were incubated with 20 µg of UV-inactivated DBs of TR-∆GFP (DB-PC+) or DBs of TR-BAC (DB-PC−) for 24 h and 48 h. Cell extracts were analyzed for the MX1, IFIT3, and ISG15 expression. (a) Western blots were probed with antibodies against MX1, IFIT3, and ISG15. The viral IE1-protein was visualized to verify an infectious virus. Internalization of DBs into endothelial cells was visualized by the detection of the phosphoprotein 65 (pp65). Tubulin was probed as the sample loading control. For quantification in (b–d), the ratio of the fluorescence intensity of each IRG protein band to the corresponding tubulin band was measured using the LICOR Image Studio Lite (Ver 5.2.5) software. The values of three independent experiments were plotted. Error bars represent the SD. Comparisons between the groups were calculated using the unpaired t-test with Welch´s correction. ** p < 0.01.

Article Snippet: Endothelial cells (ECs, HEC-LTTs; [43,44]), kindly provided by Christian Sinzger (Institute for Virology, Ulm University Medical Center, Ulm, Germany), were cultured in endothelial cell growth medium (ECGM Kit, C22110, Promocell, Heidelberg, Germany) or endothelial growth medium (EGM BulletKit; Lonza Sales Ltd., Basel, Switzerland), supplemented with 2 μg/mL doxycycline (Sigma-Aldrich, Saint Louis, MO, USA) in culture vessels coated with 0.1% gelatin (Sigma-Aldrich, Saint Louis, MO; PMID: 26345505).

Techniques: Western Blot, Expressing, Incubation, Virus, Control, Software