(A–B) Few particles are detected in AFM images of cryo-extracted samples from the buffer alone and from AR10Q cell lysates treated for 8hrs or 120hrs with DHT.
(C) AR112Q cells were lysed after 8hrs in DHT, at a time that precedes the formation of significant nuclear inclusions. Lysates were resolved by SDS-AGE and top of the gel, containing slow-migrating species, was subject to cryo-extraction and imaging by AFM. Large, amorphous particles were detected on the mica.
(D) AR112Q cells were lysed after 120hrs in DHT, when considerable nuclear inclusions have formed. Lysates were resolved by SDS-AGE and the bottom of the gel, containing fast-migrating species, was subject to cryo-extraction and imaging by AFM. The detected particles were smaller and amorphous.
(E–F) Quantification of numbers of particles per unit area for slow- and fast- migrating species.
(G) Correlation analysis between slow- and fast-migrating species demonstrates distinct and non-overlapping populations.