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. 2006 Jan;17(1):251–262. doi: 10.1091/mbc.E05-10-0936

Figure 2.

Figure 2.

Overexpression of the C-terminal domain causes loss of Scd5p cortical localization, but actin and endocytosis are normal. (A) Localization of GFP-Scd5p in GST-CT-overexpressing cells. GFP-SCD5 cells transformed with pTB338 [GAL1:GST] (SL5151) or pTMN2 [GAL1:GST-CT](SL5143) were grown to log phase in–leucine/2% raffinose medium with 0.5 μg/ml DAPI to log phase. Then cultures were induced on galactose at 30°C for 3.5 h and examined by fluorescence microscopy. (B) Localization of actin and GFP-Scd5p in GST-CT-overexpressing cells. GFP-SCD5 cells transformed with pTMN2 [GAL1:GST-CT](SL5143) were induced on galactose at 30°C for 3.5 h and then fixed and stained for simultaneous actin (Alexa-594 phalloidin) and GFP visualization. (C) Endocytosis: LY uptake in SL5151 (GST) or SL5143 (GST-CT) was performed after 3 h galactose induction (left panel). 35S-α-factor internalization was performed in wild-type cells transformed with pTB338/GST (SL5231) or pTMN2/GST-CT (SL5232) after 4-h induction on galactose (right panel).