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. 2019 Jan 23;9:3325. doi: 10.3389/fmicb.2018.03325

FIGURE 6.

FIGURE 6

Asprellcosides B inhibits influenza A virus HA-mediated hemolysis. (A) 25 μL influenza A/FM/1/47(H1N1) virus was incubated at room temperature with a twofold gradient of diluted Asprellcosides B for 30 min. Added with dilutions of 100 μL at different time-points, the virus was subsequently incubated at room temperature for 48 h. The blank control was a PBS solution. Note the inhibition on hemagglutinin of Asprellcosides B. (B) Hemolysis inhibition assay of Asprellcosides B against the influenza A/FM/1/47(H1N1) virus strain. Asprellcosides B was diluted by PBS and was mixed in a 96-well-plate with an equal volume of the influenza virus A/FM/1/47(H1N1). 1% suspension of chicken erythrocytes was added after incubation at room temperature for 30 min. The plates were incubated for 20 min at 37°C. Then, sodium acetate at different pH values (4.6, 4.8, 5.0, 5.2, 5.4, 5.6, 5.8, and 6.0) was added and incubated at 37°C for 30 min to bring about hemolysis. At the end of incubation, the plates were centrifuged at 12,000 rpm for 6 min, and the supernatants were detected. A microtiter plate reader was used to read the OD540 nm. n = 3, each pH value in triplicate. (C) The inhibition ratios (%) of hemolysis of influenza virus H1N1 A/FM/1/47 with different pH values by Asprellcosides B.