Researcher builds new model to examine Ushersyndrome NatureComms
. After fixation the eyes were transferred to 30% sucrose, embedded in OCT medium, and stored at −80 °C until they were completely frozen then four micrometer-thick sections were cut. After 10 min fixation with acetone at −20 °C, the sections were dried for 10 min. Rehydration was done in 1x PBS, followed by 30 min in blocking solution . The sections were then incubated overnight at 4 °C with the primary antibodies diluted in a blocking solution.
. The insoluble interphotoreceptor cell matrix was then separated from the soluble IPM by centrifugation at 750×for 5 min. The supernatant was considered to contain soluble components of the IPM. The pellet was washed three times in 1x PBS, resuspended in 0.1x PBS with 2x protease inhibitors and homogenized. The suspension was centrifuged at 50,000×for 30 min in a Sorvall Discovery M150 ultracentrifuge equipped with a fixed angle rotor .
. Secondary antibodies against mouse and rabbit conjugated to HRP diluted 1:15,000 in 5% non-fat dry milk were used.Eyes were orientated via marking of the superior hemisphere along the vertical meridian at the limbus with a hot needle after euthanasia. A slit in the superior cornea was made and the eyes were then fixed for 2 h in 0.1 M sodium phosphate buffer containing 2.5% glutaraldehyde, 2% paraformaldehyde, and 0.025% CaClat 4 °C.
Belgique Dernières Nouvelles, Belgique Actualités
Similar News:Vous pouvez également lire des articles d'actualité similaires à celui-ci que nous avons collectés auprès d'autres sources d'information.
New fund launched to bring nature back to life in NIOver three quarters of Northern Irish folks are worried about the impact nature loss will have on their lives, according to a YouGov survey
Lire la suite »
Circadian clock molecule REV-ERBα regulates lung fibrotic progression through collagen stabilization - Nature CommunicationsThe molecular clock REV-ERBα regulates lung injury during fibrosis, but the role of REV-ERBα in fibrogenesis remains unknown. Here, the authors show that REV-ERBα interacts with the lysyl oxidase-collagen axis during fibrogenesis and demonstrate the therapeutic potential of Rev-erbα agonist against lung fibrosis.
Lire la suite »
Type I interferon shapes brain distribution and tropism of tick-borne flavivirus - Nature CommunicationsHere the authors combine a multimodal imaging-snRNAseq transcriptomics strategy to provide insight into the distribution of a neurotropic tick-borne flavivirus in the brain, and show that absence of interferon signaling increases infection of resident microglia.
Lire la suite »
ST3GAL1 and βII-spectrin pathways control CAR T cell migration to target tumors - Nature ImmunologyCAR T cell success requires targeting tumors, but these cells can get trapped in other tissues, such as in the lungs, where they can cause pathology. Here, the authors use a loss-of-function CRISPR screen to identify regulators of CAR T cell tumor trafficking and engineer CAR T cells accordingly to overcome this limitation.
Lire la suite »
Cholinergic deficits selectively boost cortical intratelencephalic control of striatum in male Huntington’s disease model mice - Nature CommunicationsThe corticostriatal dysfunction underlying Huntington’s disease remains incompletely understood. Here, the authors find increased intratelencephalic connectivity resulting from deficient cholinergic transmission in a mouse model of Huntington’s disease.
Lire la suite »