Fig. 2.

Validation of significant sgRNA candidates (a) Expression of selected sgRNA transcript sequences in engineered T24 SAM cells was analyzed by PCR using primer listed in Additional file 6: Table S1b. b Detection of mRNA expression level of the 8 candidate genes relative to the NTC (c) Percentage of cells in S-phase 24 h after Palbociclib (1000 nM) treatment normalized to vehicle-treated control (*, P < 0.05; unpaired t-test compared percentage of cells in S-phase under Palbociclib treatment of cells to NTC). d SRB cell proliferation assay after 4 days of Palbociclib (1000 nM) treatment of cells transduced with sgRNA candidates normalized to NTC (*, P < 0.05; unpaired t-test comparing normalized absorbance of cells treated with Palbociclib or NTC). e Images and quantification of colonies of a 10-day clonogenic assay under Palbociclib (1000 nM) treatment. (Surviving fraction = colony number/seeded cell number in %; * means P < 0.05; unpaired t-test). Data represent the mean ± SD of 3 replicates