Figure 5.

[A] Deletion of polyamine transporter enhances the uptake of S. pneumoniae TIGR4 by murine alveolar macrophage (AMJ2.C8) cells. Cells were cultured in appropriate medium to 90% confluency, and were infected with TIGR4 or ΔpotABCD at MOI of 1:10 for 2 h, followed by 1 h incubation with penicillin and gentamicin at 37 °C in 5% CO₂. Cells were lysed in 0.0125% Triton X-100 for CFU enumeration. [B] Deletion of polyamine transporter enhances the invasion of S. pneumoniae TIGR4 in human BEAS.2B lung epithelial cells. Cells were cultured in appropriate medium to 90% confluency, and were infected with TIGR4 or ΔpotABCD at MOI of 1:10 for 2 h, followed by 1 h incubation with penicillin and gentamicin at 37 °C in 5% CO₂. Cells were lysed in 0.0125% Triton X-100 for CFU enumeration. [C,D] Deletion of polyamine transporter does not affect the ability of S. pneumoniae. TIGR4 to adhere to murine AMJ2.C8 [C] and human BEAS.2B [D] lung epithelial cells. Cells were cultured as mentioned in 5A, infected with TIGR4 or ΔpotABCD at MOI of 1:10 for 2 h at 37 °C in 5% CO₂, rinsed 3X in sterile PBS, and CFU enumerated. All assays were performed at least three times with two or more technical replicates. One-way ANOVA and Tukey’s multiple comparison tests were used to calculate the p-values. (****p-value = <0.0001, ns = p-value >0.05).