Journal: bioRxiv
Article Title: IFT20 regulates lymphatic endothelial cell-cell junctions via endocytic trafficking of VE-cadherin
doi: 10.1101/2025.01.15.631989
Figure Lengend Snippet: (A) Control and IFT20 KD HDLECs in homeostasis were immunostained for IFT20 and RAB5. Micrographs are MIPs from laser scanning confocal microscopy. Scale bars = 25 μm. (B) Quantification of integrated intensity from RAB5+ area in control and IFT20 KD HDLECs. Intensities are graphed relative to average control HDLEC RAB5 intensity. Quantification is from one of three biological replicates and is representative of 300+ control and 300+ IFT20 KD HDLECs. (C) Control and IFT20 KD HDLECs were serum starved in media containing 1/5 EGM-MV2 and 4/5 EBM-2 (hereafter, starving media) for 24 h. Cells were then either: placed in fresh starving media for 45 min, then fixed; placed in fresh starving media + 2 μg/mL VEGF-C for 45 min, then fixed; or placed in fresh starving media + 2 μg/mL VEGF-C for 45 min, then placed in EBM-2 basal media for a 90 min washout, then fixed. RAB5 was detected in fixed cells by immunofluorescence. Micrographs are MIPs from laser scanning confocal microscopy. Scale bar = 25 μm. (D) Quantification of C . Integrated intensities from RAB5+ area are graphed for control and IFT20 KD HDLECs separately and combined. Intensities are graphed relative to average control HDLEC RAB5 intensity in the 45 min starve condition. Data points represent the average of three FOVs within one technical replicate. Each of three independent experiments included two or three technical replicates (three FOVs each) per experimental condition. Quantification is representative of 300+ control and 300+ IFT20 KD HDLECs. *p<0.05, **p<0.005, ***p<0.001.
Article Snippet: HDLECs were routinely cultured in Endothelial Cell Growth Media MV2 (EGM-MV2) (PromoCell, C-39226) at 37°C and 5% CO 2 .
Techniques: Control, Confocal Microscopy, Immunofluorescence
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