Fig. 2

Multiplexed HCR RNA-FISH in wildtype Arabidopsis inflorescence. a 2-channel FISH for CLV3 (green) and WUS (red) in inflorescence (top-view with maximum intensity projection). HCR hairpin amplifiers B2-AlexaFlour488 (B2-AF488) and B3-AlexaFlour546 (B3-AF546) were used in the amplification stage. The arrow and arrowhead indicate the earliest flower primordia that express CLV3 and WUS respectively. The orthogonal views across the dash line were shown in the bottom. b Test for background. All FISH steps were same as those in Fig. 2a except that mScarletI-B2 and mEGFP-B3 were used in the hybridization step as negative controls. The orthogonal views across the dash line were shown in the bottom. c 3D projection of the sample in panel a. d FISH for CLV3 (green) and WUS (red) in 11-day-old shoot apical meristem (side-view). e 3-channel FISH for AG (green), AP3 (yellow), and STM (red) in inflorescence (top-view with maximum intensity projection). HCR hairpin amplifiers B1-AF546, B2-AF488, and B3-AF514 were used in the amplification stage. White arrows indicate the lateral and medial sepal primordia. White squares represent the STM expression domain between adjacent sepal primordia. Nuclei were stained by DAPI (blue). Scale bar = 100 µm