Anti-DYKDDDDK polyclonal antibody [FITC]

Cardiac ankyrin repeat protein (CARP) is a nuclear transcriptional co-factor that has additional functions in the myoplasm as a component of the muscle sarcomere. Previous studies have demonstrated increased expression of CARP in cardiovascular diseases, however, its role in cardiomyocyte apoptosis is unclear and controversial. In the present study, we investigated possible roles of CARP in hypoxia/reoxygenation (H/R)-induced cardiomyocyte apoptosis and the underlying mechanisms. Neonatal mouse ventricular cardiomyocytes were isolated and infected with adenovirus encoding Flag-tagged CARP (Ad-CARP) and lentivirus encoding CARP targeted shRNA (sh-CARP), respectively. Cardiomyocyte apoptosis induced by exposure to H/R conditions was evaluated by TUNEL staining and western blot analysis of cleaved caspase-3. The results showed that H/R-induced apoptosis was significantly decreased in Ad-CARP cardiomyocytes and increased in sh-CARP cardiomyocytes, suggesting a protective anti-apoptosis role for CARP. Interestingly, over-expressed CARP was mainly distributed in the nucleus, consistent with its role in regulating transcriptional activity. qPCR analysis showed that Bcl-2 transcripts were significantly increased in Ad-CARP cardiomyocytes. ChIP and co-IP assays confirmed the binding of CARP to the Bcl-2 promoter through interaction with transcription factor GATA4. Collectively, our results suggest that CARP can protect against H/R induced cardiomyocyte apoptosis, possibly through increasing anti-apoptosis Bcl-2 gene expression. (C) 2016 Elsevier B.V. All rights reserved.

Crop plants accumulate a large amount of storage starch and storage proteins in the endosperm. Genes involved in the biosynthesis of these substances work in concert during development of the rice endosperm. The rice flo2 mutant produces aberrant seeds with reduced grain quality. FLOURRY ENDOSPERM 2 (FLO2), the causative gene of the flo2 mutant, is considered to be a regulatory protein that controls the biosynthesis of seed storage substances. FLO2 contains tetratricopeptide repeat (TPR) motifs that may mediate protein-protein interactions. In this study, we identified the protein that interacts with the TPR motif of FLO2. We generated a transformant that produced the FLAG-tagged fusion FLO2 protein in the flo2 mutant and used this in the shotgun proteomic analysis. A protein, which we named FLOC1, interacted with FLO2. In vitro pull-down assays indicated that the TPR motif was involved in this interaction. A knock-down transformant of FLOC1 showed significantly reducted fertility and generation of seeds with abnormal features. These findings suggest that FLOC1 is involved not only in seed fertility but also in seed quality. These phenotypes were also observed on the RNAi transformants of the flo2 mutant although the effect of the flo2 mutation remained. these findings imply that there is a difference in the functions of FLO2 and FLOC1 although both of appear to be involved in the control of seed quality during seed formation.