SerRSAA exhibits stronger activity than SerRSWT in suppressing angiogenesis in mice.
(A) Western blot to measure the expression of SerRS proteins in mouse 3B11 endothelial cells stably transfected with tag-free mouse SerRSWT or mutants (SerRSAA and SerRSDD). The SerRS levels are quantified by the density of the bands and indicated relative to the level of the endogenous SerRS (Vector). (B, C) Matrigel plug angiogenesis assay performed with stably transfected 3B11 cells in C3H/HeJ mice. Matrigel plugs (dash lines enclosed regions) excised 14 days after implantation were analyzed by immunofluorescence for Vegfa (B) and quantified (C) (means ± SEM, n = 6 individual plugs per group, *p < 0.05, **p < 0.01, ***p < 0.001, n.s., Student t test). See S1 Data for quantitative data and statistical analysis. See S2 Data for original, uncropped images supporting blots and gel results. n.s., not significant; SerRS, seryl-tRNA synthetase; SerRSAA, S101A/S241A; SerRSDD, S101D/S241D; SerRSWT, wild-type SerRS; Vegfa, vascular endothelial growth factor A
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