Figure 5.

Pdpn regulates the expression of Filamin C. (A–D) Volcano plots showing differential gene expression in groups of wild type (WT), Pdpn-knockout (KO) and Pdpn-transgenic (TG) cells before and after H. pylori infection at MOI 10 for 24 h (as indicated by +Hp). Comparison was made between the group with a high Pdpn expression versus the group without or with a lower Pdpn expression, as follows: (A) Pdpn-TG versus Pdpn-KO; (B) Pdpn-TG versus WT; (C) Activated Pdpn-TG versus activated KO; and (D) Activated WT versus activated Pdpn-KO. Note that a gene Filamin C (FlnC) was repetitively shown in all four groups of comparison, indicating its consistent upregulation by high Pdpn expression. Vertically broken lines demarcate log₂ (fold change) at >1.5 or <−1.5. Horizontally broken lines demarcate statistical significance (−log₁₀ p-value > 2, or p < 0.01) by R-based program in nSolver Advanced Analysis 2.0 software. Significantly upregulated and downregulated genes were highlighted in red and blue dots, respectively. (E) Bar charts show normalized RNA transcript counts of Filamin C from NanoString analysis. Data are shown as mean ± SD of biological duplicate samples. Statistical significance by unpaired Student’s t-test (*p ≤ 0.05; **p ≤ 0.01). (F) Immunoblot analysis of Filamin C Cell lysate from the uninfected or H. pylori (MOI 10, 24 h)-infected WT, Pdpn-KO, and Pdpn-TG cells were examined. Membranes were probed with antibodies to Filamin C, Pdpn, and β-actin as loading control. Data shown are representative data of two independent experiments.