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Fig. 4 Comparison of EE and IF obtained for each target by microinjection of Alt-R HDR components and PE components. Zebrafish embryos at the one-cell stage were injected with 200 pg (col1a2 c.918A>T), 400 pg (col1a2 c.1416A>C, col1a2 c.2093G>A) or 800 pg (atp6v1e1b c.634_636CGA>TGG, atp6v1e1b c.383T>C, brca2 c.6803G>A) Cas9. EE and IF were compared with injection of PE components (RNP mixture containing 480 ng/µL pegRNA and 1,380 ng/µL PE2-His protein) into the yolk of one-cell-stage zebrafish embryos. Left-hand y axes show EE (blue) and right-hand y axes show IF (gray). The bars and error bars represent the mean + s.e.m. of three biologically independent replicates (n = 3), where each replicate represents a pool of 20–30 injected embryos. Unpaired two-tailed t-test: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.