Figure 2.

Altered CRT expression in ER-enriched vesicles from transgenic cell lines. ER-enriched vesicles from CRT, GFP transgenic, and wild-type cell lines were separated on Suc gradients. Fraction 2, containing ER-enriched membrane vesicles (Fig. 1), was analyzed by 10% (w/v) SDS-PAGE (15 μg protein lane⁻¹), blotted, and immunostained with polyclonal antibodies against maize CRT (1:5,000). A, Increased production of CRT in CRT sense lines. Lane 1, Non-heat-shocked wild type. Lane 2, Heat-shocked wild type. Lane 3, Non-heat-shocked CRT sense line (Nt CRT:7). Lane 4, Heat-shocked CRT sense line (Nt CRT:7). B, Lowered production of CRT in CRT antisense lines. Lane 1, Non-heat-shocked wild type. Lane 2, Heat-shocked wild type. Lane 3, Non-heat-shocked antisense CRT (Nt CRT-A:3). Lane 4, Heat-shocked antisense CRT (Nt CRT-A:3). C, No significant changes in CRT production in mgfp5 transgenic cell lines (Nt GFP:4). Lane 1, Non-heat-shocked wild type. Lane 2, Non-heat-shocked Nt GFP:4 transgenic line. Lane 3, Heat-shocked wild type. Lane 4, Heat-shocked Nt GFP:4 transgenic line. D, Expression of a maize CRT in the transformed line Nt CRT:7. Lane 1, Purified maize CRT. Lane 2, Heat-shocked Nt CRT:7. Lane 3, Non-heat-shocked wild type. Lane 4, Standard. E, Protein visualized with silver staining. Lane 1, Purified maize CRT. Lane 2, Heat-shocked Nt CRT:7. Lane 3, Non-heat-shocked wild type. Lane 4, Standard.