To confirm Stx phage transfer by bAGPs, we randomly chose transductant colonies from the co-culture experiment described earlier (Figure 3) for further verification with PCR to detect genes specific for the Stx2 phage (stx2), E.
We calculated the transduction frequencies by dividing the number of transductants by the number of recipients.
Pink colonies growing on sorbitol-MacConkey agar supplemented with either kanamycin and tetracycline or kanamycin and ampicillin were regarded as presumptive transductants (i.e., recipients of the stx2-encoding phage 933W).
Generation of OSC19 Transductant Lines and Cell Culture Conditions
The generation of stable UMSCC1 control and precursor miR375-expressing (Pre375) lentiviral transductant cell lines was previously described.
(45) For the matrix degradation assay, 1 X [10.sup.5] UMSCC1 transductant cells or 0.5 X [10.sup.5] OSC19 transductant cells were plated in DMEM culture media on Alexa Fluor 405 gelatin-coated MatTek dishes and cultured for 4 hours.
UMSCC1 transductant cells were lysed in radioimmunoprecipitation assay buffer containing 1X protease inhibitors (Cat.
The numbers of primary clones and transductant clones amplified were 2 x [10.sup.9] and 2.2 x [10.sup.12], respectively.
The numbers of primary clones and transductant clones amplified were 2 x [10.sup.8] and 3.2 x [10.sup.11], respectively.