Fig. 2. Expression of EPMH reporters in bloodstream trypanosomes. (A) Control untransformed bloodstream cells (BS221) and cell lines expressing EPMH or EPMHgpi were preincubated (1 hour) with (+) or without (-) tunicamycin, and then metabolically radiolabeled with [³⁵S]Met/Cys (1 hour) in the continued absence or presence of inhibitor. Reporters and endogenous VSG were immunoprecipitated from cell and media fractions with anti-EP₉ (top, 2x10⁷ cell equivalents/lane) and anti-VSG221 (bottom, 2x10⁵ cell equivalents/lane) antibodies. Mobilities of the precursor (P), and mature (M) forms of EPMH, of EPHMgpi (), and of VSG (V), are indicated on the left; mobilities of molecular mass markers on the right. (B) EPMH cells were radiolabeled with [³⁵S]Met/Cys for 2 hours and then immunoprecipitated with anti-EP₉ (lane 1). Identical anti-EP₉ immunoprecipitates were solubilized and reprecipitated with either anti-EP₉ (lane 2) or biotinyl-tomato lectin (lane 3). All lanes contain 2x10⁷ cell equivalents/lane. (C) EPMH cells were radiolabeled with [³H]Myristate for 4 hours and labeled polypeptides were immunoprecipitated from cell and media fractions with anti-EP₉ (5x10⁷ cell equivalents/lane) or anti-VSG221 (1x10⁶ cell equivalents/lane). All samples were analyzed by SDS-PAGE and phosphorimaging (panels A and B) or fluorography (panel C).