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. 2013 Mar 26;8(3):e59560. doi: 10.1371/journal.pone.0059560

Figure 2. Biological relevance of hcpG during H. pylori growth and infection.

Figure 2

(a) RT-PCR amplification of hcpG transcripts from 24 h growth in pure broth cultures. recA, positive control; nctl, negative control, in which extracted RNA was used as a template for RT-PCR. The geographic origins of the strains used were as follows: HpJ99, United States; HpG27MA, Italy; HpR10, South Africa; HpK17, South Korea; HpPeCan32, Lima, Peru; and HpS46, Spain. (b) RT-PCR amplification of hcpG transcripts in cultured AGS cells 6 h after infection (multiplicity of infection [MOI] = 100). gapdh was included as a positive control for AGS cell transcription in addition to the controls listed in Fig. 3a. UI, uninfected. (c). Quantitative RT-PCR analysis of hcpG transcripts in cultured AGS cells 3 and 6 h after infection with HpG27MA and HpJ99, respectively. The results show the averages from three experiments (± standard error of the mean). (d) Growth curves of WT HpG27MA and hcpG and/or hcpC derivatives in pure broth cultures; the results show averages from three experiments (± standard deviation [SD]). (e). Growth curves of WT HpG27MA strain and hcpG and/or hcpC deletion derivatives during infection of cultured AGS cells. The results show the averages from five experiments (± SD). (f). Verification of HcpG synthesis during infection of cultured AGS cells with the HpG27MA derivative expressing the HcpG::6xHis fusion protein; Lane 1, uninfected AGS cells (control); lanes 2 and 3, 3 h after infection with HpG27MA::HcpG-6XHis and HpG27MAΔhcpG, respectively; lanes 4 and 5∶6 h after infection with HpG27MA::HcpG-6XHis and HpG27MAΔhcpG, respectively. α-Tubulin was used as a measure for equal gel loading. CFU, colony-forming unit.