Fig. 3

YULINK interacted with GLUT1 and participated in PDGF-triggered glucose uptake and glycolysis. The bar graphs illustrate A glucose uptake and B pyruvate production in PASMCs with YULINK KD or OE treated with or without PDGF (20 ng/ml) for 24 h. The values represent the mean of three independent experiments ± standard deviation. *p < 0.05 compared to scramble control without PDGF. ^#p < 0.05 between compared groups. C Western blot analysis indicates the expression of GLUT1 and HK-2 in normal PASMCs, PASMCs with YULINK KD, and PASMCs with YULINK OE treated with or without PDGF for 24 h. The numbers labeled below the respective blot lanes indicate the relative fold normalized with the internal control. D Representative images from immunofluorescence analysis indicate the expression of YULINK (green) and GLUT1 (red), and DAPI (blue) indicates nuclear staining. Magnification 200×. E Whole-cell lysates and F membrane fractions derived from PASMCs or PAH-PASMCs with or without 20 ng/ml PDGF treatment and PAH-PASMCs were subjected to Western blot analysis for YULINK expression. β-Actin and Na/K ATPase served as internal controls. G Membrane protein lysates obtained from PASMCs subjected to indicated treatments were used for immunoprecipitation with YULINK antibody-conjugated beads. The proteins pulled down were subsequently analyzed via Western blot to assess the expression of YULINK and GLUT1. While Input serves as positive control, Isotype IgG serves as negative control