Fig. 2

FQI1 treatment diminished expression of mitotic regulators. a Schematic of experimental protocol. FQI1 or vehicle was added to HeLa cells during synchronization to the G1/S border using a double thymidine block. Cells were released from the block, including addition of 20 μM of thymidine, for subsequent analyses. b Lysates from cells treated with vehicle or 1.8 μM FQI1 were harvested at release from the G1/S block (0 h) or when control cells visually reached mitosis (~ 8 h post release) and analyzed for AURKB or CDC20 RNA levels, as normalized to levels of GAPDH RNA. Data points and means are plotted relative to the expression from vehicle treated cells at each time point and are derived from 2 to 4 independent experiments. **p = 0.0045; ****p < 0.0001. c Representative immunoblots for the indicated proteins in cell lysates harvested when control cells visually reached mitosis (~ 8 h post release from a G1/S block), after treatment with increasing concentrations of FQI1 from 0 to 3.6 μM. Molecular weight markers are indicated on the right side. Relative intensities can only be compared within each separate immunoblot of each protein showing levels at increasing FQI1 concentrations, not between separate immunoblots. The images were cropped to indicate the proteins of interest; full images are in Additional file 4. d Independent quantitation of the protein levels from cell lysates harvested for mitotic expression (as in panel c). Individual protein levels were normalized to those of β-actin from the same lysate. Data points and means are from 2 to 5 independent experiments. *p = 0.037 (FQI1 = 1.8 μM), 0.015 (FQI1 = 3.6 μM); **p = 0.0048; ***p = 0.0005 (unpaired T test)