Fig. 3. Expression patterns of miR164 miRNAs in wild-type plants. (A-D) Confocal images of inflorescences. Each transgenic plant expresses the GFP variant 3xVENUS-N7 (green/yellow) under control of the individual miRNA regulatory sequences (as indicated). In A,B, FM4-64 dye was used to stain plasma membranes (red); in C,D, organ outlines are highlighted by red chlorophyll autofluorescence. T1 plants were examined and representative expression patterns are shown. The number of plants showing depicted expression pattern (x) with respect to total sample size (n_tot), indicated as ratio (x/n_tot), was 7/7 (A), 2^*/20 (B) and 5/5 (C,D). ^*No expression was detected in 18 out of 20 transgenic lines harboring pMIR164b::3xVENUS-N7. (E-P) In situ hybridization analysis of miR164 miRNA distribution (E,F,I-P) using DIG-labeled LNA-ath-miR164a antisense oligos, in Ler wild-type (E,F), in mir164a-4 b-1 double-mutant (I,J), in mir164a-4 c-1 double-mutant (K,L), in mir164b-1 c-1 double-mutant (M,N) and in mir164abc triple-mutant (O,P) plants. The inset in K shows mir164 accumulation in developing flowers. (G,H) No signal above background was detected when DIG-labeled Scramble-miR LNA-oligo was used as a control probe on Ler wild-type tissue. Scale bars: 100 µm.