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. 2022 Feb 22;13:832931. doi: 10.3389/fphar.2022.832931

FIGURE 2.

FIGURE 2

Optimization of infection conditions for the Bac-Mam virus. Conditions for the Bac-Mam virus to infect COS-1 cells were optimized by monitoring CYP3A4 expression. CYP3A4 expression in COS-1 homogenates (20 μg) was determined by immunoblotting with Supersomes™ contained 1 pmol CYP3A4 (BD Gentest) as a standard. Viral infection was independently conducted twice in each experiment. (A) Optimization of viral amount (MOI) ranging from 0 to 150. Culture time after infection and concentration of sodium butyrate were fixed at 24 h and 3 mM, respectively. (B) The result of quantification of CYP3A4 in the optimization of viral amount. Each determined CYP3A4 level (N = 2), and the mean ± S.E.M. are presented. (C) Optimization of culture time after infection ranging from 24 to 72 h. Viral amount and concentration of sodium butyrate were fixed at MOI = 100 and 3 mM, respectively. (D) The result of quantification of CYP3A4 in the optimization of culture time. Each determined CYP3A4 level (N = 2), and the mean ± S.E.M. are presented. (E) Optimization of the concentration of sodium butyrate ranging from 0 to 4.5 mM. Viral amount and culture time after infection were fixed at MOI = 100 and 48 h, respectively. (F) The result of quantification of CYP3A4 in the optimization of concentration of sodium butyrate. Each determined CYP3A4 level (N = 2), and the mean ± S.E.M. are presented. ND, not detected.