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. 2003 Mar;77(5):3181–3190. doi: 10.1128/JVI.77.5.3181-3190.2003

FIG. 3.

FIG. 3.

Colony-forming abilities of H77 subgenomic RNAs containing mutations in NS3. (A) Huh-7.5 cells were electroporated with 0.5 μg (each) of the subgenomic replicons H/SG-Neo (L+I), H/SG-Neo (D+I), Con1/SG-Neo (I), and H/SG-Neo (pol−). Forty-eight hours later, the cells were subjected to G418 selection, and the resulting colonies were fixed and stained with crystal violet. Representative dishes after 2.5 × 104 cells were plated are illustrated. The percentage below each dish refers to the calculated G418 transduction efficiency of the replicon that was determined by serially titrating transfected cells from 2 × 105 to 1 × 103 cells per 100-mm-diameter dish, together with feeder cells electroporated with the pol− replicon. The resulting G418-resistant foci were counted for at least three cell densities, and the relative G418 transduction efficiency was expressed as a percentage after dividing the number of colonies by the number of electroporated Huh-7.5 cells initially plated. (B) One microgram (each) of the subgenomic RNAs H/SG-Neo (L+I), H/SG-Neo (L), H/SG-Neo (D), and H/SG-Neo (pol−) was transfected into Huh-7.5 cells, and after 3 weeks of G418 selection, the transduction efficiency was determined as described for panel A. Dishes seeded with 2 × 105 electroporated cells are depicted, with the relative transduction efficiencies shown below.