Skip to main content
. 2018 Apr 4;9:677. doi: 10.3389/fimmu.2018.00677

Figure 1.

Figure 1

Activity analysis of the recombinant Rv0888NS, D438A, and H481N proteins. (A) Western blotting analysis of the expression of the Rv0888NS, D438A, and H481N proteins. (B) SDS-PAGE analysis of affinity-purified recombinant Rv0888NS, D438A, and H481N proteins. (C) Cell fractionation experiments were performed to determine the subcellular localization of Rv0888NS, D438A, and H481N; KatG protein served as a cytoplasmic marker for Mycobacterium smegmatis. CL represents whole-cell lysate proteins; CW represents cell wall; CM represents cell membrane; CP represents cytoplasm; and CF represents culture filtrate. (D) Assessment of DNase activity of the recombinant Rv0888NS, D438A, and H481N proteins. (E) Enzymatic sphingomyelinase assay using purified proteins; 20 µg of purified Rv0888NS, D438A, and H481N were analyzed using the Amplex Red Sphingomyelinase Kit. Sphingomyelin hydrolysis was determined using an excitation wavelength of 530 nm, and emission was detected at 590 nm. The error bars show the SEM of three independent experiments.