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Fig. 6 Impact oflmnb1 expression on PHA-like Cells and AML development. (A) CRISPR/Cas9 targeted exon 7 of zebrafish lmnb1. The target sequence is in red, and the PAM sequence in green. The lmnb1 (?4/?4) line has deleted nucleotides, leading to a truncated protein missing the lamin tail domain. (B) lmnb1 mRNA expression in 3 dpf lmnb1 mutants and siblings, detected by qPCR. (C) PHA-like cells in 3-mo-old lmnb1 (?4/?4) zebrafish (blue arrows, green arrows, and red arrows indicate segmented, band/kidney-shaped and round nucleus neutrophil, respectively). (D) Comparation of neutrophils with different shape of nucleus in wt and lmnb1 (?4/?4). The black asterisks indicate statistical difference. (E) KM blood cell morphology in 3-mo-old c-mybhyper;pu.1G242D/G242D and c-mybhyper;pu.1G242D/G242D;lmnb1OE (blue arrows, green arrows, and red arrows indicate segmented, band/kidney-shaped and round nucleus neutrophil, respectively). (F) Comparation of neutrophils with different shape of nucleus in c-mybhyper;pu.1G242D/G242D and c-mybhyper;pu.1G242D/G242D;lmnb1OE. (G) Three 6-mo-old lmnb1KO F0 exhibited an AML-like phenotype in KM (black triangles indicate precursors, AML-like F0 zebrafish were numbered as #1, #2, and #3). (H) Calculation of different blood cell types in 6-mo-old wildtype and lmnb1KO F0 zebrafish. (I) The expression of lmnb1 in 3 AML-like lmnb1KO F0 KM blood cells was detected by qPCR. (J) Blasts increased in 3-mo-old lmnb1 (?4/?4) zebrafish (green arrows, red arrows, black triangles, and green triangles indicate lymphocytes, myelomonocytes, blasts, and precursors, respectively). (K) Calculation of different blood cell types in 3-mo-old wildtype and lmnb1 (?4/?4) zebrafish. (L) Calculation of different blood cell types in 3-mo-old c-mybhyper;pu.1G242D/G242D and c-mybhyper;pu.1G242D/G242D;lmnb1OE.