Fig. 3

Overexpression of HSPA9B-tagged proteins in L. donovani promastigotes. Protein expression in transgenic promastigotes carrying the episomal construction GFP-HSPA9B-pXG (a) or the integrative version HSPA9B-mCherry (b) were analyzed by flow cytometry and western blot. Histograms indicate fluorescence intensity of controls (black line) and transfected parasites (green or red filled). Total (CE) and immunopurified (IP) protein extracts (c) or just total (d) extracts obtained from logarithmic cultures were resolved by SDS-PAGE and probed with anti-GFP or anti-mCherry antibodies. Blots were stripped and re-probed with an anti-α-tubulin antibody as a loading control. Molecular weights are indicated in kDa