MCM4 expression in Leishmania.
A. Schematic representation of the conserved domains in LdMCM4. PIP box - PCNA interacting protein motif; A – Walker A motif; B – Walker B motif; R – arginine finger; Z – zinc finger; S1 – sensor 1; S2 – sensor 2. B. SDS-PAGE (10% PAGE) analysis of overexpressed and purified recombinant LdMCM4 (2 ug; ∼97.2 kDa). C. Western blot analysis of recombinant LdMCM4 with mouse anti-MCM4 antibody (1∶5000 dilution). D. Western blot analysis (10% SDS-PAGE) using anti-MCM4 antibodies (dilution of 1∶1000). First panel - lane 1, recombinant MCM4; lane 2, L. donovani whole cell extracts. Second panel - Whole cell extracts from Leishmania promastigotes and amastigotes (4×107 cell equivalents) probed with anti-MCM4 antibodies and anti-tubulin antibodies (Zymed Laboratories; 1∶2000 dilution; loading control). E. Western blot analysis (10% SDS-PAGE) of whole cell extracts from Leishmania procyclics and metacyclics (4×107 cell equivalents) probed with anti-MCM4 antibodies and anti-tubulin antibodies (loading control).
