Culturing L. mexicana Promastigotes and Evaluating Macrophage Infection

The L. mexicana wild type promastigotes were cultured in the M199 medium (Sigma-Aldrich) containing 10% FBS supplemented with 1% Basal Medium Eagle vitamins (Sigma-Aldrich), 2% sterile urine, and 250 µg/mL Amikacin (Bristol-Myers Squibb, New York, NY, USA), and 200 µg/mL of Hygromycin B (Carl Roth GmbH) for BnSP-7-ecDHFR-HA mutant cell lines. Infection of the J774 macrophages was performed as described previously [49 (link),52 (link)]. In addition, infection was evaluated by counting Giemsa-stained slides as described previously [51 (link)]. All experiments were performed with two independent biological replicates and samples were analyzed in triplicate.