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. 2018 Jan 31;13(1):e0191570. doi: 10.1371/journal.pone.0191570

Fig 2. Optimizing transduction conditions for the A673 target cell line.

Fig 2

Effects of (A) polybrene and (B) centrifugal spin force on target cell transduction. A673 and HEK293 cells were transduced with 100 μl of non-silencing shRNA lentivirus in a total volume of 2 ml containing 10% FBS. Transduced cell fractions were determined by flow cytometry analysis of GFP expression 72 h later. (C) Influence of transduction volume and serum content was evaluated using 100 μl of LaminA/C lentivirus in spin transductions with 8 μg / ml of polybrene. (D) Effects of viral packaging using distinct ratios of shRNA and packaging plasmids. GAPDH shRNA was packaged in HEK293T cells as indicated and A673 target cells were infected with 100 μl of these viruses using centrifugal spin and 8 μg / ml of polybrene in 2 ml volume containing 10% FBS. Asterisk (*) indicates manufacturer’s recommendation; (**) indicates ratio employed in (A)-(C). All graphs represent mean ±standard deviation (SD) of three independent experiments. Student’s t-test did not reach significance (p < 0.05).