Extended Data Fig. 9: Migrasomes rescue VEGFA/CXCL12 KD induced capillarization and monocyte recruitment defects.
From: Monocytes deposit migrasomes to promote embryonic angiogenesis
(a) CAMs were treated with VEGFA siRNA. 72 h after transfection, the CAMs were harvested, and the VEGFA knockdown efficiency in the CAMs was determined using q–PCR. Data are presented as mean ± SEM; n = 16 pieces from three independent experiments; P values were calculated using a two-tailed, unpaired t-test, P < 0.0001. (b) CAMs treated with CXCL12 siRNA for 72 h were harvested, and CXCL12 knockdown efficiency in CAM pieces was detected by q–PCR. Data are presented as mean ± SEM; n = 16 pieces from three independent experiments; P values were calculated using a two-tailed, unpaired t-test, P < 0.0001. (c) VEGFA and CXCL12 knockdown efficiencies in siRNA-treated CAM pieces were determined by western blot analysis using then indicated antibodies. (d) CAMs were transfected with the indicated siRNAs. After 48 h, migrasomes embedded in low-melting-point agarose were added. After another 48 h, CAMs were visualized by stereomicroscopy. Scale bar, 1 mm. (e) CAMs treated with VEGF siRNA (top) or CXCL12 siRNA (bottom) from (d) were quantified for the change in the number of sprouting capillaries. Data are presented as mean ± SEM; n = 9 fields from three independent experiments; in siVEGFA group, P values were calculated using a two way ANOVA unpaired multiple comparisons, N.S: no significance (P=0.7626, 96h/0h, siNC+Migrasome vs. siVEGFA+Migrasome); P = 0.0003(96h/0h, siNC+Agarose vs. siNC+Migrasome); P < 0.0001; in siCXCL12 group, P values were calculated using a two way ANOVA unpaired multiple comparisons, P=0.0028 (96h/0h, siNC+Migrasome vs. siCXCL12+Migrasome); P=0.0012 (96h/0h, siCXCL12+Agarose vs. siCXCL12+Migrasome); P<0.0001. Source numerical data and unprocessed blots are available in source data.
