Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Antiangiogenic Effects and Therapeutic Targets of Azadirachta indica Leaf Extract in Endothelial Cells

doi: 10.1155/2012/303019

Figure Lengend Snippet: (b)

Article Snippet: DPP4 , 15.39 ± 4.21 , 15.78 ± 2.62 , Hs00175218_m1 , Aminopeptidase.

Techniques: Binding Assay, Protein Binding, Activity Assay, Ubiquitin Proteomics

Journal: Viruses

Article Title: Unveiling the Role of TMPRSS2 in the Proteolytic Activation of Pandemic and Zoonotic Influenza Viruses and Coronaviruses in Human Airway Cells

doi: 10.3390/v16111798

Figure Lengend Snippet: Activation of SARS-CoV and MERS-CoV S by TMPRSS2 and further host proteases. ( A , B ) HeLa cells were co-transfected with plasmids coding for MERS-CoV S ( A ) or SARS-CoV ( B ) with a C-terminal FLAG-tag and plasmids coding for human TMPRSS2 or HAT or murine TMPRSS4 or TMPRSS13. Cell lysates were analysed via SDS-PAGE at 48 h p.t. Western blot analysis was performed using an antibody targeting the C-terminal FLAG-tag. Tubulin was used as a loading control. The data shown are representative of three individual experiments. EV = empty vector. ( C , E ) HeLa cells co-expressing MERS-CoV ( C ) or SARS-CoV ( E ) S, as well as the respective receptors DPP4 or ACE2, were incubated for 48 h. After fixation, the cells were stained with a primary antibody targeting the C-terminal FLAG-tag of S and a fluorescence-coupled secondary antibody. DAPI was used to stain the nuclei. Representative images of three independent experiments are shown. The scale bar represents 100 µM. ( D , F ) Quantification of MERS-CoV S ( D ) or SARS-CoV ( F ) mediated cell–cell fusion. For each condition, ten randomly taken images were analysed by counting the nuclei per syncytium (with at least three nuclei). The data shown are the mean values + SEM of three independent experiments. Statistical significance was determined with a one-way ANOVA followed by Šídák’s multiple comparisons test. ns = not significant, ** = p < 0.01, **** = p < 0.0001.

Article Snippet: The expression vectors coding for SARS- or MERS-CoV S with a C-terminal FLAG-tag, pCMV3-MERS-Spike-cFLAG (VG40069-CF) and pCMV3-SARS-Spike-cFLAG (VG40150-CF), as well as a vector encoding human dipeptidyl peptidase 4 (DPP4) with a C-terminal HA-tag, pCMV3-hDPP4-cHA (HG10688-CY), were purchased from Sino Biological, Houston, TX, USA.

Techniques: Activation Assay, Transfection, FLAG-tag, SDS Page, Western Blot, Control, Plasmid Preparation, Expressing, Incubation, Staining, Fluorescence