(A) Representative microscopy images showing CL-11 bound to B16 cells. CL-11 (red) and DAPI (blue). Scale bar: 10 μm. (B) Representative microscopy images of EdU staining in B16 cells following the treatment with rCL-11 at the indicated concentrations. EdU (green) and DAPI (blue). Scale bar: 200 μm. (C) Cell proliferation rate and total cell numbers corresponding to the images in B. B and C are representative of 2 independent experiments. Data are expressed as mean ± SEM. (D and E) Additional sets of EdU cell proliferation assay performed using 1,200 ng/mL of rCL-11. (D) Representative microscopy images of EdU staining. Scale bar: 100 μm. (E) Cell proliferation rate and total cell numbers. Data were analyzed by paired t test (5 individual experiments). Each dot represents the average value of the percentages or cell numbers calculated from 5 image fields at 100× for each sample. (F) Left panel: Western blots showing total and phosphorylated ERK (p-ERK), JNK (p-JNK), and AKT (p-AKT) in B16 melanoma cells at the indicated time points following rCL-11 (600 ng/mL) stimulation. Right panel: quantification of phosphorylated proteins relative to their respective total proteins corresponding with the blots. (G) Representative Western blots showing total and p-ERK, p-JNK, and p-AKT in B16 melanoma cells that have been stimulated with rCL-11 at the indicated concentrations for 5 minutes. (H) Quantification of p-ERK, p-JNK, and p-AKT relative to their respective total proteins corresponding to the blots in G. (I) Representative Western blots showing total, p-EGFR, and p-HER3 in B16 cells following the treatment with rCL-11 for 5 minutes. (J) Quantification of p-EGFR and p-HER3 relative to their respective total proteins corresponding with the blots in I. (H and J) Data were analyzed by 1-way ANOVA (5 independent experiments). *P < 0.05; **P < 0.01.