Journal: Nature

Article Title: GlycoRNA complexed with heparan sulfate regulates VEGF-A signalling

doi: 10.1038/s41586-025-10052-8

Figure Lengend Snippet: a , Quantification of 10E4, Siglec-11, 9D5, cs-DDX21 and cs-hnRNP-U intensity per cell from three independent staining experiments on HUVECs. Data are mean ± s.e.m. a.u., arbitrary units. b , Western blot analysis of whole-cell lysates isolated from HUVECs after starvation and treatment with an RNase pool followed by 3 ng ml −1 VEGF-A 165 , VEGF-A 121 or EGF stimulation (top). Quantification of the ratio of pERK to total ERK was also calculated across the biological triplicates (bottom). Statistical assessment was performed with a two-sided Student’s t -test and P values are shown. Data are mean ± s.e.m. c , Representative images of starved HUVECs treated with an RNase pool, and subsequently with VEGF-A 165 or VEGF-A 121 , finally stained with anti-VEGF-A 165 (red) or anti-VEGF-A (red). Statistical assessment was performed with a Student’s t -test and P values are shown. Data are mean ± s.e.m. NS, not significant. d , Representative images of HUVECs treated with an RNase pool and stained with anti-VEGFR2 (red). Statistical assessment was performed with a Student’s t -test and P values are shown. Data are mean ± s.e.m. e , Representative images of starved HUVECs treated with VEGF-A 165 and then co-stained with anti-VEGF-A 165 (purple) and Siglec-11 (yellow). Three independent experiments were performed. f , Schematic of the microfluidic chip (top left) used to grow HUVECs without (top middle) and with (top right) RNase A for 6 days. Representative images of BFP expressed in the HUVECs. Statistical assessment of the total migration area was performed using an unpaired two-sided Student’s t -test (bottom). Data are mean ± s.e.m. Four independent experiments were performed. g , Representative image (maximum z -projection view (left) and z -projection slice view (right)) of sprouts from a +RNase A device for the cells (blue), F-actin (red) and PECAM1 (green). Four independent experiments were performed.

Article Snippet: For VEGF-A 165 , VEGF-A 121 and VEGFR2 staining, 1 μg ml −1 of anti-VEGF-A 165 (R&D Systems), 1 μg ml −1 of anti-VEGF-A (Proteintech), and 1 μg ml −1 anti-VEGFR2 (R&D Systems) were precomplexed with 0.5 μg ml −1 of goat anti-rabbit AF647 secondary antibody and donkey anti-goat AF647 secondary antibody (Thermo Fisher Scientific).

Techniques: Staining, Western Blot, Isolation, Migration