Anti-LAT monoclonal antibody, clone MBU-2E2

Loss of chondroitin sulfate (CS) has been reported to play a key role during intervertebral disc degeneration (IDD). However, the detailed mechanism of CS and its synthases have not been elucidated. Since CS is mainly synthesized by chondroitin synthases 3 (Chsy3), here, the Chsy3 knockout mice are generated by using CRISPR-Cas9 and semi-cloning technology to study its mechanism during IDD. We find that CS and Chsy3 expression are decreased during IDD both in human and mice nucleus pulposus (NP) tissue, and knockout of Chsy3 shows that spontaneous IDD phenotype resembles that of human samples in theChsy3(-/-)mice. Taking advantage of RNA-Seq data, we confirm increased catabolic and decreased anabolic changes inChsy3(-/-)NP cells. By using bioinformatic analysis and validation, we find that Hippo signaling pathway is significantly downregulated, and the activation of Yap1 is mainly affected inChsy3(-/-)NP cells. Furthermore, functional analyses have shown that Chsy3 could regulate NP cell degeneration by Actin tension mediated activation of Yap1, which is independent of Hippo/Lats signaling. In summary, our findings reveal a novel mechanism that depletion of CS-related Chsy3 can cause spontaneous intervertebral disc degeneration by mediating Yap activation through CS-related actin-tension in NP cells.

Precise control of the crystal orientation and hierarchical crystalline morphology is essential for application of crystalline polymers, although a homogeneous morphology with a controlled crystal orientation is rarely formed in polymer crystallization. Here we report the formation of a homogeneous surface morphology with submicrometer-scale periodicity by simple crystallization of polymer brushes on a solid substrate in poor solvents. Unlike the complex morphology commonly observed for crystallization in air, a periodic line pattern spontaneously formed when a thin brush layer of poly(ecaprolactone) (PCL) was isothermally crystallized in marginally poor solvents. The length scale of the periodic line pattern was similar to 100 nm in pitch and a few nanometers in height. The structure was found to depend on the crystallization temperature, brush layer thickness, and affinity of the poor solvent to the polymer. Grazing-incidence X-ray scattering experiments revealed that the PCL crystals within the periodic line pattern were predominantly aligned in the f lat-on orientation. A structure formation mechanism based on the kinetic balance between the secondary nucleation and lateral growth processes at the growth front of crystal lamellae was proposed. The poor solvents acted as a plasticizer that enhanced the mobility of brush chains and modulated the nucleation and growth rates, which led to a highly ordered morphology with little branching.