Mycobacterium tuberculosis Rv0431 expressed in Mycobacterium smegmatis, a potentially mannosylated protein, mediated the immune evasion of RAW 264.7 macrophages

Prediction of glycosylation sites and analysis of mutant Rv0431 proteins and Rv0431 protein. A. Prediction of glycosylation sites. 59Thr, 60Thr, 61Thr, 62Thr were predicted to be glycosylated with possibility. B. SDS-PAGE of purified mutant Rv0431 proteins and purified Rv0431 protein in ΔM5447. M. PageRuler Prestained Protein Ladder (Fermentas); Lane 1. Rv0431m3/M; lane 2. Rv0431 expressed in ΔM5447; lane 3. Rv0431m1/M; lane 4. Rv0431m2/M; lane 5. Rv0431m4/M; lane 6. Rv0431/M. C. Western blot of purified mutant Rv0431 protein and Rv0431 protein in ΔM5447 using anti-His tag antibody. M. PageRuler Prestained Protein Ladder (Fermentas); Lane 1. Rv0431m3/M; lane 2. Rv0431 expressed in ΔM5447; lane 3. Rv0431m1/M; lane 4. Rv0431m2/M; lane 5. Rv0431m4/M; lane 6. Rv0431/M. D. Western blot of purified Rv0431 protein and Rv0431 protein in ΔM5447 using ConA. M. PageRuler Prestained Protein Ladder (Fermentas); Lane 1. Rv0431m3/M; lane 2. Rv0431 expressed in ΔM5447; lane 3. Rv0431m1/M; lane 4. Rv0431m2/M; lane 5. Rv0431m4/M; lane 6. Rv0431/M.140