Figure 3

Knockdown of P4HA2 suppresses breast cancer cells invasiveness in 3D culture. (A) Phase images of control and P4HA2-silenced MDA-MB-157 and MDA-MB-231 cells in 3D culture for 4 days. The right column shows immunofluorescence images of F-actin (red) and DAPI (blue) staining. P4HA2-silenced MDA-MB-157 and MDA-MB-231 cells formed smaller and less invasive cell clusters than control cells (scale bar, 50 μm). (B) Quantification of the invasive branches of control and P4HA2-silenced MDA-MB-157 and MDA-MB-231 cells in 3D culture by counting the branches in at least 50 colonies. Knockdown of P4HA2 reduced the invasive branches. (C) EdU-staining was used to analyze the proliferation of control and P4HA2 knockdown MDA-MB-231 cells in 3D culture. Knockdown of P4HA2 had little effect on the proliferation of MDA-MB-231 cells (**p < 0.01). (D) Transwell cell invasion assay analysis of control and P4HA2-silenced MDA-MB-231 cells (scale bar, 200 μm). (E) Quantification of invasion analysis of control and P4HA2-silenced MDA-MB-231 cells. Knockdown P4HA2 significantly inhibited cell invasion in MDA-MB-231 cells compared with control group.