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. 2012 Nov 8;7(6):922–931. doi: 10.1111/irv.12035

Figure 2.

Figure 2

 Experimental design of protocols for testing effect of the alkaloid compounds by HA assay at different time points. Virus mixtures contained 0·01 MOI. 1, negative control; 2, virus only as positive control; 3, mixture of virus at 0·01 MOI and different compounds (AA1:0·52 μm; AA2:82·07 μm; AA3:4·15 μm; AA4:13·45 μm, Oseltamivir: 0·625 μm, 100 μl) were incubated at 4°C for 1 h, inoculated onto cells 1 h and then the media were replaced with drug‐free DMEM for 72 h; 4, same as 3, except the replacement media DMEM contained compounds; 5, after adsorption 1 h, the culture containing Amaryllidaceae alkaloids compounds; 6, mixture of virus at 0·01 MOI and Amaryllidaceae alkaloids compounds were added to cells at 37°C for 1 h, and then replaced with drug‐free DMEM for 72 h; 7, same as 6, except the replacement media contained drugs. Supernatants from the above seven protocols were collected at different time points (12, 24, 36, 48, 60 and 72 h) to test the HA titer.