Journal: bioRxiv
Article Title: Kindlin-2-Moesin interaction orchestrates sprouting angiogenesis via modulating endothelial membrane mechanics and VEGF signaling
doi: 10.64898/2026.02.24.707842
Figure Lengend Snippet: (A) Schematic illustration of tamoxifen administration in pregnant mice. (B) Left: representative images of E12.5 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) embryos. Middle and right: representative confocal images of E12.5 embryonic brain sections stained with IsoB4 (red). (C) Quantification of vascular coverage in the brain shown in (D) (n = 7/5 embryos). (D) Schematic of tamoxifen administration schedule in Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) mice. Tamoxifen was administered from P1 to P3, and retinas were analyzed at P6. (E) Quantification of Fermt2 mRNA (encode Kindlin-2) levels in brain endothelial cells (ECs) isolated from P6 Kindlin-2 iΔEC(Pdgfb) pups (n = 3/3 pups). (F) Representative confocal images of flat-mounted retinas from P6 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups stained with IsoB4 (red). (G-H) Quantification of (H), showing reduced vascular area and branch points in Kindlin-2 iΔEC(Pdgfb) pups (n = 9/9 pups). (I) Representative images of the angiogenic front in P6 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) retinas stained with IsoB4. Higher-magnification images show blunted morphology of tip cells. (J) Quantification of (K), showing a reduced number of sprouts in Kindlin-2 iΔEC(Pdgfb) pups (n = 4/4 pups). (K) Representative confocal images of ERG (EC nuclei, green) and IsoB4 (red) in flat-mounted retinas from P6 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (L) Quantification of ERG + nuclei in (M) (n = 5/5 pups). (M) Representative confocal images of Ki67 (green) and IsoB4 (red) in flat-mounted retinas from P6 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (N) Quantification of Ki67 + cells in the vascular area of Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups (n = 9/8 pups). (O) Representative images of brains from P6 Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups and immunofluorescence staining of Ter119 (erythrocytes, green), IsoB4 (red) in brain sections. White arrows indicate hemorrhage area. (P-Q) Quantification of (Q), showing reduced vascular coverage and branch points in Kindlin-2 iΔEC(Pdgfb) brains (n = 5/6 pups). Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 by two-tailed Student’s t-test. Scale bars: 1 mm in brightfield panels of (B) and (O); 500 μm in (B), (F), and low magnification of (K), and (M); 200 μm in low magnification of (I), and high magnification of (K), and (M); 100 μm in (O) and high magnification of (I).
Article Snippet: Vibratome sections were washed, permeabilized in PBS containing 0.3% Triton X-100, blocked in PBS containing 2% BSA and 0.3% Triton X-100 for 1 hr, and incubated overnight at 4 °C with Ter119 (R&D Systems, #MAB1125, 1:200).
Techniques: Staining, Isolation, Immunofluorescence, Two Tailed Test
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