Journal: Nature Cell Biology

Article Title: Haematopoietic stem and progenitor cell heterogeneity is inherited from the embryonic endothelium

doi: 10.1038/s41556-023-01187-9

Figure Lengend Snippet: a , UMAP of defined EHT cluster cells from kdr + ECs in the tail of WT and 128 Δ/Δ at 26 hpf. b , UMAP representing efnb2a , gata2b and cmyb expression normalized with Z -score. c , UMAP of gata1a , hmgn2 and lcp1 expression normalized with Z -score, in nHSPC cmyb + clusters (C3, C6, C8 and C5). d , RNA velocity trajectories in nHSPC clusters showing that C8.nHSPC pLEPs and C5.nHSPC pLMPs are terminal states of C3.nHSPCs and C6.nHSPCs. e , Violin plot of lymphoid ( ikzf1 , P > 0.9999; hmgb2a , P = 0.0001; hmgn2 , P < 0.0001), erythroid ( gata1a , P < 0.0001; alas2 , P < 0.0001; cahz , P < 0.0001) and myeloid ( lcp1 , P < 0.0001; spi1b , P < 0.0001; cebpa , P < 0.0001) markers in WT cells of C8.nHPSC pLEPs and C5.nHSPC pLMPs. Statistics represent the comparison between C8.nHSPC pLEPs and C5.nHSPC pLMPs for each gene (ordinary one-way ANOVA). f , UMAP cell cycle analysis on nHSPC clusters. g , Quantification of S, G2/M and G1 phase in C3. and C6.nHSPCs. C3.nHSPCs cells are mainly in S phase and G1, while C6. nHSPCs in G2/M (two-tailed Student’s t -test with Bonferroni post-hoc correction). h , Confocal images of IF using anti-RFP, anti-GFP and EdU staining ( n = 22 (WT) and 20 (128 Δ/Δ ) embryos; P = 0.0032) or anti-pH3 ( n = 18 (WT) and 24 (128 Δ/Δ ) embryos; P = 0.0208) in Tg( kdrl:mCherry s896 ,cmyb:GFP zf169 ) AGM at 32 hpf. S phase and G2/M nHSPCs are increased ( kdrl +, cmyb +, EdU+ or pH3+ blue arrowheads and kdrl +, cmyb +, EdU− or pH3− white arrowheads) in miR-128 Δ/Δ (three independent experiments; two-tailed Mann–Whitney test). i , Violin plot of gata1a ( P = 0.0004) , ikzf1 ( P = 0.0006 and 0.0015) and lcp1 ( P = 0.4505 and 0.5383 ) expression in clusters C8.nHPSC pLEPs and C5.nHSPC pLMPs per genotype (Mann–Whitney test). j , k , Model of nHSPC heterogeneity acquired during EHT in the AGM at 26 hpf WT ( j ) and 128 Δ/Δ ( k ). 128 Δ/Δ nHSPC heterogeneity is biased towards S and G2/M nHSPCs (green circles), and erythroid and lymphoid primed nHSPCs (blue circles, bigger size represents increase gene expression but not number). Not signifcant (NS): P > 0.05. ** P ≤ 0.01, *** P ≤ 0.001. LP, lymphatic progenitor; ISVs, intersegmental vessels; DA, dorsal aorta; PCV, posterior cardinal vein.

Article Snippet: The antibodies used are: mouse anti KDR-PE (0.25 µg per 10 6 cells, Biotechne, cat. no. MAB3572 clone 89106), mouse anti CD34-PE-Cy7 (BD, cat. no. 348791 clone 8G12), mouse anti CD43-FITC (BD, cat. no. 555475 clone 1G10), mouse anti CD73-PE (BD, cat. no. 550257 clone AD2), mouse anti CXCR4-APC (BD, cat. no. 555976 clone 12G5) and mouse anti CD235a-APC (BD, cat. no. 551336 clone HIR-2) (refs. , , ).

Techniques: Expressing, Comparison, Cell Cycle Assay, Two Tailed Test, Staining, MANN-WHITNEY

Journal: bioRxiv

Article Title: Inhibition of peripheral VEGF signaling rapidly reduces leucocyte obstructions in brain capillaries and increases cortical blood flow in an Alzheimer’s disease mouse model

doi: 10.1101/2021.03.05.433976

Figure Lengend Snippet: ELISA measurements of VEGF1R receptor ( A ), VEGF2R receptor ( B ) from brain lysates, and soluble VEGF1R receptor ( C ), and soluble VEGF2R receptor ( D ) from plasma samples after 1 week of anti-VEGF-A treatment or saline injections in APP/PS1 and WT mice (n=3 mice per group). Kruskal-Wallis test with multiple comparison correction to compare across groups. In all graphs each point represents the ELISA measurement from one mouse, red horizontal line represents the median; * indicates p = 0.05.

Article Snippet: After the measuring the concentrations ELISAs were performed following the manufacture’s protocol (VEGF1R (MVR100) and VEGF2R (MVR200B), R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Saline, Comparison