Journal: Antimicrobial Agents and Chemotherapy

Article Title: Cepharanthine inhibits enterovirus entry by endolysosomal deacidification and exhibits protective activity in vivo

doi: 10.1128/aac.00764-25

Figure Lengend Snippet: BBAs delayed EV71 entry through the endosomal compartment. ( A ) HeLa cells were inoculated with EV71 stock at an MOI of 300 at 4°C for 1 h for adsorption. Afterward, the cells were treated with TET or CEP at 10 µM for 90 min before being fixed and permeabilized. Puncta signals (green) representing EV71 virions were detected using a rabbit anti-VP2 Ab, followed by detection with a secondary Ab conjugated to Alexa Fluor 488. DAPI was used to stain the nucleus (blue). Scale bar represents 20 µM. ( B ) The number of puncta surrounding each cell was calculated based on images from five fields. ( C ) Fluorescence signals were detected at the indicated time points after adsorption with EV71. Cells were co-stained with Ab against viral VP2 and ( D ) EEA1 (indicating early endosomes) or ( E ) LAMP1 (indicating lysosomes). The portion of VP2 colocalized with EEA1 or LAMP1 is shown as the average ± SEM for more than 200 cells. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: Additionally, cells were co-stained with either EEA1 Ab Alexa Fluor 546 (G-4, 1:500, sc-137130, Santa Cruz) or LAMP1 Ab Alexa Fluor 546 (H4A3, 1:500, sc-20011, Santa Cruz).

Techniques: Adsorption, Staining, Fluorescence