Figures & data
Figure 1A-C. (A) Sodium dodecyl sulfate PAGE (SDS–PAGE) analysis of M. tuberculosis Mtb9.9 protein purification. Lane 1, molecular weight markers; lane 2, purified Mtb9.9A protein; lane 3, purified Mtb9.9C protein; lane 4, purified Mtb9.9D protein; lane 5, purified Mtb9.9E protein. Proteins were visualized with Coomassie brilliant blue staining. (B) Mass-finger printing analysis of the purified Mtb9.9A by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF). (C) Mass-finger printing analysis of the purified Mtb9.9C by MALDI-TOF.
Figure 2. Analysis of antigen-specific production of interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-4 by mice immunized with purified recombinant Mtb9.9 proteins in IFA and PBS in IFA, respectively. Splenocytes were then co-cultured with purified recombinant Mtb9.9 proteins or PBS as control. Data are represented as mean ± standard deviations (n = 4). *p < 0.05, **p < 0.01, ***p < 0.001. 1、3、5、7、9、11、13、15: PBS-induced; 2&10: Mtb9.9A-induced: 4&12: Mtb9.9C-induced; 6&14: Mtb9.9D-induced; 8&16: Mtb9.9E-induced.
Figure 3. Serological responses to M. tuberculosis Mtb9.9 family proteins in immunized mice. Serum samples were collected 3 weeks after the last injection and analyzed by enzyme-linked immunosorbant assay (ELISA) for the presence of anti-Mtb9.9 family proteins IgGH+L (A); anti-Mtb 9.9A IgG1, IgG2b and IgG2c (B); anti-Mtb 9.9C IgG1, IgG2b and IgG2c (C); anti-Mtb 9.9D IgG1, IgG2b and IgG2c (D); and anti-Mtb 9.9E IgG1, IgG2b and IgG2c (E).
Figure 4. Mtb9.9 proteins underwent western blot analyses for Mtb9.9A (A), Mtb9.9C (B), Mtb9.9D (C), and Mtb9.9E (D).
Table 1. Polymerase chain reaction (PCR) primers and thermal cycle parameters for amplification of Mycobacterium tuberculosis open reading frames
Figure 1D-E. (D) Mass-finger printing analysis of the purified Mtb9.9D by MALDI-TOF. (E) Mass-finger printing analysis of the purified Mtb9.9E by MALDI-TOF. The spectrum indicates the molecular mass of the recombinant proteins (N-terminal 6 × His-tag) were 13.185181, 13.174290, 13.078204 and 13.078379 kDa respectively.
