Project description:To identify the developmentally regulated genes, which could confound identification of PN and CQ drug responsive genes, RNA samples from drug-free synchronized cultures from ring, trophozoite, and schizont stages were individually labelled and hybridized with a pooled sample from the three stages. The data from this experiment were used to compare the developmental profile of the K1 strain with the data from other P. falciparum strains. 9 samples were obtained from three developmental stages of parasite development (3 each from ring, trophozoite and schizont synchronized parasites). Three independent cultures were obtained, synchronized on different days.
Project description:The urgent need to address water scarcity underscores the importance of enhancing plant drought resistance. This study investigates whether pretreatment with abscisic acid (ABA) activates early stress signaling, thereby improving barley drought response when subsequently exposed to drought conditions. Although the individual responses to drought and ABA are well-documented, their synergistic effects in barley warrant further investigation. This study examines the impact of ABA on barley drought resilience through an experimental design that incorporates four distinct treatments: optimal watering, ABA application at 60 days post-sowing, and two drought stress treatments - one with and the other without prior ABA application. Key physiological parameters, such as photosynthesis, stomatal conductance and chlorophyll content, were analyzed in conjunction with transcriptomics. The results suggest that ABA pretreatment initiates early stomatal closure and elevates the expression of essential genes like NCED1, BG8, and HvA22, priming barley for improved drought resistance. During the drought, ABA-pre-treated barley maintained high chlorophyll levels, indicating sustained photosynthetic activity, a trend that persisted across treatments during the post-drought recovery phase. Furthermore, ABA pre-treatment was found to preserve photosystem II efficiency during drought conditions. Transcriptomic analyses revealed distinct gene expression profiles, alternative splicing profile and isoform switching, highlighting the molecular complexities of ABA role in drought response. These alterations span stress response, metabolic pathways, and DNA modification processes, providing a comprehensive view of ABA treatment's regulatory and metabolic impacts. In conclusion, ABA pretreatment strengthens barley drought defense by fostering stomatal closure and gene activation, guiding research strategies grounded in ABA and suggesting that genotypes with elevated ABA levels could have enhanced resilience and recovery capabilities.
Project description:We used proximity dependent biotin identification (BioID) method to determine cell cycle specific interaction partners of PCDH7. HeLa S3 cells were synchronized to interphase and mitosis using double tymidine block and double thymidine block followed by S-Trityl-L-cysteine (STC) treatment respectively. Candidate interactors were isolated using streptavidin affinity purification and identified using LC-MS/MS analysis.
Project description:Microarray data from G2-synchronized p53(+) and p53(-) fibroblasts before and after 3 h release from cell cycle blockade in the presence of 5 uM sodium arsenite. Experiment Overall Design: Cells expressing p53 from a tet-off regulated construct were synchronized in G2 with a two-step procedure using 24 h aphidicolin treatment for initial G1 synchronization and 12 h of Hoechst 33342 to effect a G2 blockade. During Hoechst treatment, tetracycline was added to suppress p53 in half the cultures. Cells were then released into media containing 5 µM sodium arsenite and the appropriate concentration of tetracycline to maintaining p53 expression. mRNAs were collected at 0 h and 3 h after release from G2 synchrony.
Project description:MiR-30e represses the osteogenic program in mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) by targeting IGF2, and drives their differentiation into adipogenic or smooth muscle lineage, respectively. In Experiment 1, SMCs were cultured from mouse aortas and transduced with miR-30e or control lentivirus. In Experiment 2, MSCs were extracted from mouse bone marrow and transduced with miR-30e or control lentivirus. In Experiment 3, the MSCs were transfected with a scrambled oligo or anti-miR-30e oligo. In all 3 experiments, N=3 per group. RNA was extracted from each experiment and run on Affymetrix arrays.
Project description:MiR-30e represses the osteogenic program in mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) by targeting IGF2, and drives their differentiation into adipogenic or smooth muscle lineage, respectively. In Experiment 1, SMCs were cultured from mouse aortas and transduced with miR-30e or control lentivirus. In Experiment 2, MSCs were extracted from mouse bone marrow and transduced with miR-30e or control lentivirus. In Experiment 3, the MSCs were transfected with a scrambled oligo or anti-miR-30e oligo. In all 3 experiments, N=3 per group. RNA was extracted from each experiment and run on Affymetrix arrays.
Project description:MiR-30e represses the osteogenic program in mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) by targeting IGF2, and drives their differentiation into adipogenic or smooth muscle lineage, respectively. In Experiment 1, SMCs were cultured from mouse aortas and transduced with miR-30e or control lentivirus. In Experiment 2, MSCs were extracted from mouse bone marrow and transduced with miR-30e or control lentivirus. In Experiment 3, the MSCs were transfected with a scrambled oligo or anti-miR-30e oligo. In all 3 experiments, N=3 per group. RNA was extracted from each experiment and run on Affymetrix arrays.
Project description:[Hela cells]: We performed cdr2 knockdown with a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [Rat1a wild type and myc null cells]: We performed cdr2 knockdown using a pool of 4 cdr2-specific siRNAs to test whether cdr2 may regulate c-myc target genes as cells passage through mitosis. [HeLa cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis [Rat-1 wild type (TGR) and c-myc null (15.19) cells]: Cells were transfected with control or cdr2 siRNAs and then cells were synchronized in mitosis using a sequential thymidine/nocodazole block. Cells were then subsequently released from nocodazole block and after 3 hours RNA was harvested for microarray analysis