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. 2022 Oct 5;11(19):3128. doi: 10.3390/cells11193128

Figure 8.

Figure 8

Syringin (SRG) blocked the interaction of PR50 and C1QBP in the yeast two-hybrid-based growth assay. (A) Schematic drawing the principle of the yeast two-hybrid-based growth assay in identifying inhibitors of PR50–C1QBP interactions. (B–E) Log-phase cultures of diploid yeast cells containing plasmids encoding either Gal4 BD-/Gal4 AD-, Gal4 BD-p53/Gal4 AD-T, Gal4 BD-PR50/Gal4 AD-C1QBP (C3) or Gal4 BD-C1QBP (C3)/Gal4 AD- PR50 were washed in water. We performed spot assays on non-selective (SD/-Leu/-Trp) and selective (SD/-Leu/-Trp/-Ade/-His) plates with serial dilutions of SRG and incubated them at 30 °C for 3 days. (F–I) Overnight cultures of diploid yeast cells containing plasmids encoding either Gal4 BD-/Gal4 AD-, Gal4 BD-p53/Gal4 AD-T, Gal4 BD-PR50/Gal4 AD-C1QBP (C3) or Gal4 BD-C1QBP (C3)/Gal4 AD-PR50 in a non-selective medium were washed in water and inoculated into non-selective and selective media at OD600 = 0.2 in triplicates to implement the optical density measures every 12 h for 2 days.