FIGURE 6.

A 96-well microtiter plate based assay probing the affect of block copolymers on cellular proliferation via FGFR3c expressing BaF3 cells. In the case of the 30-kDa block copolymers (SNS and NSN), interactions with FGF1 (A) and FGF2 (B) indicated that the block copolymers with high levels of non-reducing end sulfation, SNS, appeared to be a better promoter of cellular proliferation through the FGF-HS-FGFR ternary complex than its complement, NSN. In the case of FGF7 (C) there was no clear difference between the levels of cellular proliferation promoted by SNS against NSN. These near-zero levels of proliferation were expected from FGF7 and are consistent with previously published literature (48). When comparing the 20-kDa block copolymers (SN and NS), better levels of proliferation are seen from the block copolymer with high levels of non-reducing end sulfation (SN) versus the block copolymer with high reducing end sulfation (NS). In the cases of FGF1 (D) and FGF2 (E), there are obvious differences in the levels of proliferation, similar to those seen in panel A and B. There were low-to-zero background levels of proliferation for F, the interaction with FGF7. Each of these FGF-block copolymer-FGFR combinations was tested in 8 replicates in a 96-well plate. These data were normalized against a positive control of FGF1-heparin-FGFR3c growth and a negative control of zero GAG added.