Fig. 7. Knockdown of H2AZ causes a similar phenotype to atswc6. (A) The RNAi construct targeting all three Arabidopsis H2AZ genes - HTA8, HTA9 and HTA11 (AtH2AZs). The tandemly fused HTA8, HTA9 and HTA11 cDNAs were arranged as inverted repeats which were separated by a spliceable intron. The construct was inserted between the CaMV 35S promoter and the OCS terminator. (B) FRI and AtH2AZs RNAi transgenic plants grown for 45 days under long-day conditions. (C) Distribution of flowering time in AtH2AZs RNAi T1 transformants. Plants were grown in long-day conditions. The number of plants in each category is given above each bar. (D) Expression of HTA8, HTA9, HTA11 and FLC in three AtH2AZs RNAi lines that flowered with 35, 30 and 30 leaves. Homozygous lines were selected from the T2 generation of the three transgenic plants and RNA was extracted from 10-day-old plants grown in long-day conditions. All three AtH2AZs RNAi lines showed reduced expression of HTA8, HTA9 and HTA11. The transcript level of FLC was also reduced. (E) The relative levels of HTA8, HTA9 and HTA11 transcripts in the three AtH2AZs RNAi lines as compared with FRI wild type. (F) FRI, atswc6 FRI, amiR-HTA8, amiR-HTA9, amiR-HTA11 and amiR-HTA9&11 T1 plants grown for 40 days after germination in long-day conditions. (G) RT-PCR analysis of expression of HTA8, HTA9, HTA11 and FLC in FRI, atswc6 FRI, amiR-HTA8, amiR-HTA9, amiR-HTA11 and amiR-HTA9&11 T1 plants. RNA was extracted from leaves of T1 plants. TUB was used as an internal control.