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. 2015 Jun 12;59(7):3853–3863. doi: 10.1128/AAC.04816-14

FIG 1.

FIG 1

Modulation of LdMAPK1 expression in Leishmania parasites. (a) Agarose gel stained with ethidium bromide showing PCR products of MAPK1, NEO, and HYG genes with Dd8+/+ (wild-type parasites), Dd8+/− (parasites with MAPK1 single-allele replacement), and Dd8−/− (parasites with MAPK1 double-allele replacement). (From left) Lanes 1 and 8, 1 kb plus DNA ladders; lanes 2 to 4, MAPK1 PCR products from Dd8+/+, Dd8+/−, and Dd8−/− parasites; lanes 5 to 7, NEO PCR products from Dd8+/+, Dd8+/−, and Dd8−/− parasites; lanes 9 to 11, HYG PCR products from Dd8+/+, Dd8+/−, and Dd8−/− parasites. (b) Effect of modulation of LdMAPK1 expression on promastigote growth. (c) Changes in expression of LdMAPK1 in wild-type (Dd8+/+) parasites, vector control (Dd8Vc) parasites, overexpressing transfectants (Dd8++/++), single-allele replacement mutants (Dd8+/−), and add-back mutants (Dd8+/−/+) by Western blot analysis using anti-LdMAPK1 antibodies. α-Tubulin was used as a control. (d) Quantitative estimation of expression of LdMAPK1 in cells by measuring the relative density of each band. The data are representative of the results of 3 independent experiments. The error bars represent SD. ***, P < 0.001; ns, not significant.