Extended Data Fig. 7: Identification and characterization of POLAR, POLAR-like1 (PL1) and PL2 loss-of-function mutants. | Nature

Extended Data Fig. 7: Identification and characterization of POLAR, POLAR-like1 (PL1) and PL2 loss-of-function mutants.

From: POLAR-guided signalling complex assembly and localization drive asymmetric cell division

Extended Data Fig. 7

ac, Schematic representation of the tDNA insertions in the polar-2 (SALK_146639) (a), pl1-1 (SALK_121087) (b) and pl2-1 (SALK_123929) (c) mutants. d, Schematic representation of the POLAR and PL1 CRISPR mutants polar-3 and pl1-2, respectively, and the double CRISPR mutant polar-4 pl1-2. Guide sequences of the selected target sites of the wild-type and of the CRISPR-edited polar are presented under the scheme. In the boxes, the protospacer-adjacent motif sequences are shown in red and inserted or deleted nucleotides in green. The resulting truncated amino acid sequence of the protein is shown below the scheme. Amino acids in green correspond to the wild-type POLAR protein and those in red to random amino acids produced by the frame shift after the nucleotide insertion/deletion. e, Wild-type (Col-0), POLAR tDNA (polar-2) and POLAR CRISPR (polar-3) mutants and POLAR-like1 (PL1) tDNA (pl1-1) and PL1 CRISPR (pl1-2) mutants, as well as the corresponding double tDNA (polar-2 pl1-1) and CRISPR (polar-4 pl1-2) mutants grown for 4 weeks in soil. Scale bars, 2.5 cm. f, POLAR, PL1 and PL2 gene expression measured by qRT–PCR in seedlings of polar-2, pl1-1 and pl2-1 plants. n = 3 biologically independent experiments. Primers used are depicted in the scheme and listed in Supplementary Table 2. For e and f, experiments were repeated independently three times. Box plots show the first and third quartiles, split by the median (line) and mean (cross).

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