Project description:We studied the effects of long-term endogenous expression of the melanoma oncogene RAC1 P29S using melanocyte cultures isolated from transgenic mice
Project description:We studied the effects of acute activation of the melanoma oncogene RAC1 P29S using a tamoxifen-inducible ER-fusion protein system in mouse melanocytes
Project description:The therapeutic potential of neurotrophic factors has been hampered by their inability to achieve adequate tissue penetration. Brain blood vessels, however, could be an alternative target for neuro-salvage therapies by virtue of their close proximity to neurons. Here we show that hemizygous deletion of Rac1 in mouse endothelial cells (ECs) attenuates brain injury and edema following focal cerebral ischemia. To explore the mechanisms whereby decrease of Rac1 in ECs provide neuroprotection, ECs were derived from Rac1+/+ and Rac1+/- mice. Microarray analysis was performed to determine the differential gene expression between Rac1+/+ and Rac1+/- ECs. Rac1+/+ and Rac1+/- ECs were cultured to subconfluence. Total RNA was extracted and reverse transcribed. Rac1+/+ (n = 2) and Rac1+/- (n = 2) EC cDNAs were labeled with Cy3 and Cy5, respectively, and equimolar mixtures of labeled Rac1+/+ and Rac1+/- probes were hybridized to 2 slides (sample #21648, #21649). Additionally, Rac1+/+ (n = 2) and Rac1+/- (n=2) EC cDNAs labeled with Cy5 and Cy3 (dye-swapping) were hybridized to 2 slides (sample #21650, #21651). Comparison of signal intensities between Rac1+/+ (channel 2) and Rac1+/- (channel 1) was conducted on each slide (sample), and the ratio of intensities from 4 slides were statistically assessed for each gene feature to investigate differential gene expression.
Project description:The microarray was conducted from RNA isolated from mouse mammary glands harvested from Involution day 2 mice. Cre-mediated specific Rac1 gene deletion in luminal mammary epithelial cells was achieved by crossing Rac1fx/fxRosaYFP mice with WAPiCreTg/⢠mice to produce Rac1fx/fxYFP ;WAPiCreTg/⢠(Rac1-/-) mice. Rac1fx/fxYFP mice that lacked the Cre gene were used as wild type (WT) controls. The RNA was isolated from the inguinal gland 4 for each mouse
Project description:The therapeutic potential of neurotrophic factors has been hampered by their inability to achieve adequate tissue penetration. Brain blood vessels, however, could be an alternative target for neuro-salvage therapies by virtue of their close proximity to neurons. Here we show that hemizygous deletion of Rac1 in mouse endothelial cells (ECs) attenuates brain injury and edema following focal cerebral ischemia. To explore the mechanisms whereby decrease of Rac1 in ECs provide neuroprotection, ECs were derived from Rac1+/+ and Rac1+/- mice. Microarray analysis was performed to determine the differential gene expression between Rac1+/+ and Rac1+/- ECs.
Project description:To study recombination at the fine-scale, we used high-throughput sequencing of 300 to 1,000 crossovers within the RAC1 R gene hotspot. This revealed focused intragenic crossovers, overlapping exons encoding the TIR, NBS and LRR domains. To examine the role of recombination pathways, we repeated this experiment in recq4a recq4b, fancm and recq4a recq4b fancm mutants. Finally, in order to investigate how varying patterns of interhomolog divergence influence local patterns of crossover frequency, we repeated RAC1 pollen typing sequencing in different F1 hybrids.
Project description:We report the high-throughput profiling of Tbx2 binding sites in mouse melanoma B16 cells. We generated B16 clones that have endogenous Tbx2 tagged with 3xHA, and used HA antibody for immunoprecipitation. This study provides a high-quality genome-wide Tbx2 binding profile and helps further understand Tbx2 role in melanoma progression
Project description:Wholeskin from transgenic mice overexpressing epidermal specific (K14) activated mutant (V12) Rac1 analyzed with Illumina MouseRef8 v2 Gene Expression arrays Three V12 Rac1 7 day old pups compared to three 7 day old litter-mate controls