Trypanosomes deficient in either TbAMPKβ or TbAMPKγ express GPEET-procyclin. Parasites were grown in normal SDM-79 (light grey line) or reduced glucose medium (−Glc, dark grey line), and cells induced to silence TbAMPK subunits for 7 days (+tet). Induced cells were also grown in reduced glucose medium (+tet −Glc), or in media with glucose supplemented (5 mM) (+ tet, −Glc/+Glc). Parasites (5 × 106) were fixed and stained with antibodies to EP-procyclin (TBRP1/247) or GPEET-procyclin (monoclonal 5H3). Primary antibodies were detected with FITC-conjugated anti-mouse secondary antibody and cells (10,000) analyzed by flow cytometry.