FIGURE 4.

Manual screening and combination screening of hit candidates based on secondary assay. (A) Manual screening of the significant compounds identified from the secondary hit validation assay. All samples were manually quantified in a blinded manner after 24 h treatment with candidate compounds and MTZ as described above. (n = 7 to 8; one-way ANOVA F = 16.72, p < 0.001, post-hoc Fishers LSD *p < 0.05, **p < 0.01). (B) Heatmap matrix showing the BHS for testing hit compounds in combination. All candidate compounds were 10 μM in concentration. The combination of etodolac-nepafenac, etodolac-protionamide, and etodolac-aloperine showed greater BHS compared to the administration of either alone. 0.2% DMSO for positive control and 9 mM MTZ for negative control. (n = 12 to 16; *p < 0.05, **p < 0.01, unpaired t test).