Figure 1 | High levels of knockdown achieved after induction with doxycycline.Figure 1 | pTRIPZ lentiviral vector
Figure 2 | High levels of knockdown achieved after induction with doxycycline. Knockdown of indicated target genes in HEK293T cells compared to a non-silencing control as determined by QPCR. Each bar represents an experimental duplicate while QPCR was performed triplicate using 18S rRNA as an internal reference. HEK293T cells were transduced at an MOI = 0.3, puromycin selected (5 µg/mL), and induced with doxycycline (1 µ/mL).
Figure 3 | Residual tetracycline in media does not result in leaky expression. The pTRIPZ vector does not express shRNA to GAPDH at levels capable of producing significant knockdown in the absence of doxycyline (whether in media with residual tetracycline or in special tetracycline free media).
Each bar represents four tissue culture replicates, all triplicated in QPCR for a total of 12 data points each. Statistical analysis was performed, and no statistically significant differences were found between nonsilencing and GAPDH shRNA samples. Transduction at an MOI = 5 was performed in HEK293T cells after which cells were selected with 2 µg/mL puromycin for 96 hrs.