Lack of core 1 O-glycans does not affect intracellular trafficking or turnover of PDPN in vitro. (A) Confocal images of cultured primary LECs from WT or EHC C1galt1^−/− mouse skin, with antibodies against PDPN and Tn antigen. DAPI indicates cell nuclei. Scale bar, 5 μm. (B) WT or C1galt1^−/− ECs transfected with PDPN-eGFP were treated with 100 μg/ml CHX for the indicated time. A total of 20 μg of lysates was run on a 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and probed with anti-PDPN (8.1.1) and anti-actin antibodies. The relative PDPN intensity is calculated by normalization of PDPN band intensity with actin band intensity from the same lysate. Marked line charts are presented as remainder of relative PDPN intensity at the indicated time points against that at 0 hours of treatment. The data represent the mean ± standard deviation (SD) from 4 experiments. CHX, cycloheximide.