Fig 2: Correlation between high expression of WAVE2 and poor outcome in PDAC patients. (A) Kaplan-Meier analysis of PDAC-specific survival and (B) overall survival according to WAVE2 expression. The dashed lines represent the lower and upper limits of the 95% confidence interval

Fig 3: Association of WAVE2 with ACTN4. A, Immunoprecipitation of WAVE2 from S2‐013 cells cultured on fibronectin. Rabbit IgG isotype control antibody was used as the control. Proteins within immunoprecipitates were examined on Western blots probed with anti‐WAVE2 antibody. B, Proteins in immunoprecipitates were examined with silver staining. Rabbit IgG isotype control antibody was used as the control. A 100‐kDa band is indicated by the arrow. C, The percent coverage for ACTN4 is represented by the identified peptides in the total protein sequence (accession number NP_004915). D, Immunoprecipitation of WAVE2 or ACTN4 from S2‐013 cells cultured on fibronectin. Proteins within immunoprecipitates were examined on Western blots probed with antibodies against WAVE2 and ACTN4. Rabbit IgG isotype control antibody for WAVE2 and mouse IgG isotype control antibody for ACTN4 was used as controls. E, Confocal immunofluorescence microscopic images. S2‐013 cells were cultured on fibronectin and then labeled with anti‐WAVE2 (green) and anti‐ACTN4 (red) antibodies. Arrows, WAVE2 co‐localized with ACTN4 in cell protrusions. Blue, DAPI staining. Scale bars, 10 µm

Fig 4: Signaling pathway molecules associated with WAVE2 and ACTN4. A, Human phosphoprotein arrays showing the differential phosphorylation of proteins between WAVE2 rescue construct-transfected S2-013 cells that had been transfected with both WAVE2 siRNA and ACTN4 siRNA, and WAVE2 rescue construct-transfected S2-013 cells that had been transfected with WAVE2 siRNA alone. Data are representative of three independent experiments. B, Densitometric analysis of the results of A. The level of phosphorylated p27 in WAVE2 rescue construct-transfected S2-013 cells that had been transfected with WAVE2 siRNA alone was compared to that in WAVE2 rescue construct-transfected S2-013 cells that had been transfected with both WAVE2 siRNA and ACTN4 siRNA. Data are derived from three independent experiments. Columns, mean; bars, SD. *P < 0.01 compared with WAVE2 rescue construct-transfected S2-013 cells that had been transfected with both WAVE2 siRNA and ACTN4 siRNA (Student's t test).

Fig 5: Immunohistochemistry with anti-WAVE2 antibody. A, Representative immunohistochemical staining of PDAC tissues using anti-WAVE2 antibody showing low expression of WAVE2. Arrow, the islet of Langerhans. Magnification: ×200. B and C, Representative immunohistochemical staining of PDAC tissues using anti-WAVE2 antibody showing high expression of WAVE2. Arrow, the islet of Langerhans. Magnification: ×40 (B) and ×200 (C). D, Expression of WAVE2 in the normal pancreas. Arrow, the islet of Langerhans. Magnification: ×200