Skin fibroblasts with a larger HA glycocalyx are more responsive to TGF-β
and produce more collagen when cultured in a high glucose medium. Skin fibroblasts from either wild type (WT) or Has1/3 null mice were cultured in normal glucose (1 g/L) or in high glucose (4.5 g/L) in the absence or presence of TGF-β
1 (2 ng/mL) in DMEM media with 1% FBS for 48 hours. (a) Erythrocyte exclusion assay to visualize the pericellular coat on WT and Has1/3 null fibroblasts [51
]. Cells were labeled with calcein to delineate the cell bodies. The HA glycocalyx is indicated between the white and yellow lines. (b) Western blot of protein from WT or Has1/3 null fibroblasts, probed with an antibody to type 1 collagen. GAPDH, loading control. (c) Quantification of protein band intensities from two independent western analysis experiments, performed under the conditions shown in panel (b) and normalized to GAPDH. White bars
, WT fibroblasts. Gray bars
, Has1/3 null fibroblasts. Error bars
, mean ± half-range.