gene ontology analysis Categorization of genes according to protein class was done using PANTHER Classification Systems. For each protein class, PANTHER calculates the number of genes identified in that category in both the list of dif ferentially regulated genes and a reference list contain ing all the probe sets present on the chip and compares these results using the binomial test to determine if there are more genes than expected in the differentially regulated list. Over representation is defined by p 0. 05. Functional Analysis identifying the biological func tions that were most significant to the data set were car ried out using Ingenuity Pathways Analysis. Right tailed Fishers exact test was used to calculate a p value deter mining the probability that each biological function and or disease assigned to that data set is due to chance alone.
Transfection, RNA interference and immunoblotting SiRNA against human LKLF and control siRNA was purchased from Santa Cruz Biotechnology. 4 �� 106 HMC 1 cells were transfected with 200 pmol of siRNA using Amaxa Cell Line Nucleofector Kit L with program T 020 in an Amaxa Nucleofector II device according to the manu facturers instructions. Two days after transfection, cells Anacetrapib were treated with imatinib for up to 15 h. During imatinib treatment, aliquots were prepared for analysis by TUNEL staining or immunoblot. For immunoblot analysis, whole cell lysates were pre pared using 1 �� SDS buffer, 10% glycerol, 5% beta mercaptoethanol, 0. 01% bromphenole blue.
Then, cell lysates were analyzed for cleavage fragments of caspase 3 by immunoblot analysis using a polyclonal antibody against cleaved caspase 3 or GAPDH as described previously. Knockdown of KLF2 was verified by semi quantitative RT PCR and quantitative analysis was performed using TINA2. 0 soft ware. Apomixis, asexual reproduction through seed, is wide spread among flowering plant families, but low in its fre quency of occurrence. Different from sexual reproduction, apomictically derived embryos develop autonomously from unreduced ovular cells instead of through fertilization of a reduced egg by a sperm. There fore, the progeny of an apomictic plant are genetically identical to the maternal plant. This trait can be used as an advanced breeding tool in agriculture since it would enable fixation of hybrid vigor and seed propaga tion of desirable genotypes.
No major agriculturally important crop possesses this trait. Introgression of apomixis into crops through crossing has been impeded by factors such as polyploidy and incompatibil ity. Therefore, discovery of genetic mechanisms underlying apomixis will be crucial for manipulation of apomixis for introduction into target crops. Apomixis has been classified into two types and three developmental pathways, gametophytic apomixis, including apospory and diplospory, and sporophytic apomixis, which is also known as adventitious embryony. In sporophytic apomixis, an embryo forms directly from an ovular cell and c
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