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. 2008 Jun 23;28(17):5196–5208. doi: 10.1128/MCB.00079-08

FIG. 1.

FIG. 1.

Profilin inhibits AR and Htt aggregation in cultured HEK293 cells. (A) HEK293 cells were cotransfected with constitutively active cofilin(S3A) or inactive cofilin(S3E) with Htt exon 1(Q72) CFP/YFP or ARN127(Q65) CFP/YFP. Aggregation was measured by FRET, and the effects were expressed as relative values as described in Materials and Methods. Cofilin(S3A) increased the aggregation of both Htt exon 1(Q72) and ARN127(Q65), whereas cofilin(S3E) had little effect on Htt exon 1(Q72) aggregation (**, P < 0.005 by paired t test) and decreased ARN127(Q65) aggregation. Error bars represent the standard errors of the means (SEM). (B and C) HEK293 cells were cotransfected with Htt exon 1(Q72) CFP/YFP or ARN127(65) CFP/YFP and increasing amounts of profilin-1 (which will subsequently be referred to as profilin, unless otherwise stated) (B) or profilin-2a (C). Relative aggregation was measured by FRET. Both profilin isoforms dose-dependently reduced aggregation. Error bars represent the SEM. (D) HEK293 cells were cotransfected with Htt exon 1(Q72) YFP or ARN127(Q65) YFP and profilin-1. Cells were cultured for 2 days, fixed, and analyzed using fluorescence microscopy. Profilin-1 reduced the number of inclusions formed by both Htt exon 1(Q72) YFP and ARN127(Q65) YFP. Profilin-2a had the same effect (data not shown). Representative images (magnification, ×10) are shown.