Figure 4.

HY transformation on changes in replication environment. A) Adaptation of HY to RNA-depleted NBH. 10²-fold diluted HY brain material was subjected to 11 sPMCAb rounds conducted at 10-dilution fold between rounds. The products of the 8th round were used to seed sPMCAb reactions at 100-dilution fold between rounds. End products of the 11th round are referred to as HY^R−. sPMCAb were conducted in RNA-depleted NBH. Undigested 10% NBH is shown as a reference. B) Readaptation HY^R− to NBH. HY^R− was subjected to 10 sPMCAb rounds at 100-fold dilution between rounds. sPMCAb were conducted using NBH. HY^R− readapted to NBH is referred to as HY^R+. C) Analysis of PK resistance. Western blots of sPMCAb-derived HY, HY^R− and HY^R+ treated with increasing concentrations of PK as indicated. D) Analysis of amplification rates. A series of sPMCAb reactions were seeded with 10⁴-fold diluted HY brain material (top panel), n-fold diluted HY^R− or HY^R+ (middle panels), and HY^R+ or 263K^R+ (bottom panel) and conducted using 30-, 100-, 300-, 10³-, 10⁴-, 10⁶-, or 10⁷-fold dilutions between rounds, as indicated, where n equals the dilution fold between rounds. All Western blots were stained with 3F4 antibody.