Fig. 2

Vascular niche expression of prkcda increases the number of long-term HSC clones (A) Schematic outline of the long-term fate-mapping experiment. (B) GESTALT barcodes were amplified from peripheral blood samples and quantified using SABER. The number of HSC clones at each time point is shown for sele:prkcda-2A-mCherry transgenics (prkcda) and clutchmate controls. Representative data from one of two experiments is shown. Boxes indicate mean ± SEM; each point represents a biological replicate. For 3 mpf to 22 mpf, prkcda vs. control: p = 0.0008, robust two-way mixed ANOVA. Post hoc testing for prkcda vs. control at 6 and 9 mpf: 6.5 ± 0.7 vs. 3.6 ± 0.4 HSC clones, p = 0.001 and 7.0 ± 0.8 vs. 4.2 ± 0.8 HSC clones, p = 0.02, Welch’s t test. (C) Correlation of serial samples to baseline for GESTALT barcode-identified fish. Each row per column represents a biological replicate. (D) The Pearson correlation for each fish is plotted by group and by time point (6 mpf and later). Boxes represent mean ± SEM; each point represents a biological replicate. For prkcda vs. control: mean Pearson coefficient of 0.58 ± 0.11 vs. 0.90 ± 0.03, p = 0.03, Welch’s t test. (E) Representative plots showing HSC clonal dynamics in control and prkcda groups. The height of each polygon at the indicated time points is proportional the measured frequency of alleles derived from uniquely-barcoded HSC clones. Only HSC clones with a contribution of 2% or greater are included.