Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Recombinant C-terminal proGRP18-125 stimulates proliferation and migration of DLD-1 cells. (A) Proliferation of DLD-1 cells in response to the indicated concentrations of proGRP18-125 without and with the GRP-R receptor antagonist D-Phe⁶,Leu-NHet¹³,des-Met¹⁴–bombesin(6-14) was assessed by a MTT colorimetric assay. In a typical experiment the absorbance in control wells was 0.162 ± 0.011 compared to 0.270 ± 0.008 in wells stimulated with 50 nM proGRP18-125 and 0.285 ± 0.008 in wells stimulated with 200 nM proGRP18-125. Data are the means ± SEM of at least three separate experiments each performed in quadruplicate, and are expressed as a percentage of the control OD value from wells without stimulants. (B) Migration was measured by creation of a linear wound on a confluent monolayer of serum-starved DLD-1 cells, which were then treated with the indicated concentrations of proGRP18-125 without and with the GRP-R receptor antagonist for 24 h. The width of the wound was measured at 5 distinct sites from the microphotographs taken at 17 h (gray bars) or 24 h (white bars), and expressed as a percentage of the value at 0 h (black bars). Data are the means ± SEM of at least three separate experiments. Statistical significance was assessed by one-way analysis of variance ( *, p < 0.05; **, p < 0.01; #, p < 0.001, compared to control). proGRP18-125 significantly stimulated proliferation and migration, and the stimulation was not inhibited by the GRP receptor antagonist.