Figure 6. Hypoxia promotes M2 macrophages polarization and tumorigenesis by activating ERK signaling.
RAW264.7 cells were incubated with 20μM PD in the presence or absence of IL6 under hypoxic conditions. (A) Flow cytometric analysis was performed to analyze the percentage of CD209+ cells. (B) Proteins were detected by western-blot analysis. (C) RAW264.7 cells were transfected with siRNAs specifically targeting ERK1/2 or control siRNAs, as described in the Materials and Methods. (C) Flow cytometric analysis of the percentage of CD209+ cells. (D) Proteins were detected by western-blot analysis using specific antibodies. (E) BMDMs were exposed to PD in the presence or absence of IL6. Flow cytometric analysis was used to analyze the percentages of F4/80+CD209+ cells. (F) Colocalization of p-ERK and CD209 was visualized by immunofluorescence staining (G) PD inhibits HUVEC cells tube formation. (H and I) PD inhibits LLC cells migration.