Project description:The importance of unanchored Ub in innate immunity has been shown only for a limited number of unanchored Ub-interactors. We investigated what additional cellular factors interact with unanchored Ub and whether unanchored Ub plays a broader role in innate immunity. To identify unanchored Ub-interacting factors from murine lungs, we used His-tagged recombinant poly-Ub chains as bait. These chains were mixed with lung tissue lysates and protein complexes were isolated with Ni-NTA beads. Sample elutions were subjected to mass spectrometry (LC-MSMS) analysis.
Project description:Transcriptional profiling of mouse lungs by comparing PBS and saline treated lungs with SEA and bleomycin treated lungs, respectively. Mice strains used in this study include wild type (C57BL/6), IL-13Ra1 and IL-13Ra2 deficient mice.
Project description:BCL-2 modulates IRE1a activation to attenuate ER stress and pulmonary fibrosis. Mice were treated with Bleomycin + Vehicle or Bleomycin + Navitoclax. Lungs were harvested 21 days after bleomycin injury followed by enzymatic digestion of the lungs to create single cell suspension and flow-sorting based isolation of total epithelial and stromal cells
Project description:Transcriptional profiling of mouse lungs by comparing PBS and saline treated lungs with SEA and bleomycin treated lungs, respectively. Mice strains used in this study include wild type (C57BL/6), IL-13Ra1 and IL-13Ra2 deficient mice. In mouse model of allergic asthma, mice were sensitized twice (day0 and day14) by i.p. injection of 10 µg of SEA. On days 28 and 31 mice were anesthetized with a mixture of xylazine and ketamine and given an intratracheal (I.T) airway challenge with 10 µg of SEA. Mice were sacrificed 24 h after the final airway challenge (day 32) and lungs were collected for RNA preparation. Biological replicates include 5 PBS treated and 5 SEA treated mice. In bleomycin-induced fibrosis odel, mice were anaesthetized with a xylazine and ketamine cocktail and given 0.15 U bleomycin sulfate (EMD) in saline or saline alone via the I.T. route. On day 7, mice were sacrificed and ungs were collected for RNA preparation. Biological replicates include 4 saline treated and 4 bleomycin treated mice. Goal was to determine IL-13 receptors regulated genes during SEA induced allergic asthma and bleomycin-induced fibrosis. Fluorescent cDNA targets were prepared from a 20 μg experimental RNA sample (SEA or bleomycin challenged group–dUTPCy5 – Amersham, Piscataway, NJ) and a 20 μg reference RNA sample (PBS or saline treated group–dUTPCy3- Amersham, Piscataway, NJ). Equal quantities of the above labeled cDNA (experimental and reference labeled RNA samples) were mixed and any free label present in the sample was removed by washing 3 times using a 10 kDa cutoff Vivaspin filters (Millipore). Labeled fluorescent cDNA targets were hybridized on the Whole Mouse Genome Oligo Microarray Kit (Agilent, Palo Alto, CA) containing more than 41000 gene probes.
Project description:Introgressed variants from other species can be an important source of genetic variation because they may arise rapidly, can include multiple mutations on a single haplotype, and have often been pretested by selection in the species of origin. Although introgressed alleles are generally deleterious, several studies have reported introgression as the source of adaptive alleles-including the rodenticide-resistant variant of Vkorc1 that introgressed from Mus spretus into European populations of Mus musculus domesticus. Here, we conducted bidirectional genome scans to characterize introgressed regions into one wild population of M. spretus from Spain and three wild populations of M. m. domesticus from France, Germany, and Iran. Despite the fact that these species show considerable intrinsic postzygotic reproductive isolation, introgression was observed in all individuals, including in the M. musculus reference genome (GRCm38). Mus spretus individuals had a greater proportion of introgression compared with M. m. domesticus, and within M. m. domesticus, the proportion of introgression decreased with geographic distance from the area of sympatry. Introgression was observed on all autosomes for both species, but not on the X-chromosome in M. m. domesticus, consistent with known X-linked hybrid sterility and inviability genes that have been mapped to the M. spretus X-chromosome. Tract lengths were generally short with a few outliers of up to 2.7 Mb. Interestingly, the longest introgressed tracts were in olfactory receptor regions, and introgressed tracts were significantly enriched for olfactory receptor genes in both species, suggesting that introgression may be a source of functional novelty even between species with high barriers to gene flow.