The Sla2p-binding region of LC suppresses growth and endocytic phenotypes of HC-deficient cells. (A) Growth of chc1 strain (SL2726) transformed with pRS426 (empty, 2μ), YCp50-CHC1 (CHC1, CEN), pKH2 (CLC1, 2μ), pTMN27 (clc1-[77-233], 2μ), pTMN29 (clc1-[41-233], 2μ), or pTMN28 (clc1-[1-143], 2μ). (B) Endocytosis. MATa chc1Δ bar1–1 (SL3593) was transformed with YEp24-CHC1 (CHC1, 2μ), pTMN28 (clc1-[1-143], 2μ), pKH2 (CLC1, 2μ), pRS426 (empty 2μ), or pTMN27 (clc1-[77-233], 2μ). α-factor uptake was analyzed as described in Materials and Methods. (C) Immunoblot of extracts from chc1 (SL2726) transformed with pKH2 (CLC1, 2μ), pRS316 (empty, CEN), pTMN28 (clc1-[1-143], 2μ), pTMN27 (clc1-[77-233], 2μ), or pTMN29 (clc1-[41-233], 2μ) detected with anti-Clc1p antibodies. Asterisk indicates the band corresponding to plasmid-encoded LC fragments. Note the aberrant mobility of the LC (1–143) fragment.