Figure 6
CXCR1 signaling in A549 cell lines was decreased by the depletion of REEP5 and REEP6.
(A) Using RNAs from A549 cells, RT-PCR was performed with specific CXCR1, CXCR2, and REEP1–6 primers. The PCR products were resolved on a 1% agarose gel. SM, size markers (1 Kb Plus DNA ladder from Invitrogen). (B) qRT-PCR was performed for REEP5 and REEP6 mRNA in A549 cells infected with lentivirus containing shRNAs. Relative mRNA levels of REEP5 and REEP6 were normalized with β-actin mRNA and are shown as mean ± S.E (n = 4). The bar graphs are presented by relative value to sc control. (C) A549 cells containing shRNAs were treated with IL-8 for the indicated times (min) and analyzed by western blotting with anti-pERK1/2 antibodies. NT, not treated. (D) A549 cells containing shRNAs were treated with IL-8 for 20 min, fixed, and stained with phalloidin-TRITC. The scale bar denotes 10 μm. The results are from at least three independent experiments.
