Calized to hair cell kinocilia and supporting cell primary cilia that when mutated causes non-syndromic recessive deafness in humans [67]. One of the most regularly upregulated gene in each Epha2-mutant and Epha2-null lenses was that for WD-repeat and FYVE-domain-containing protein-1 (WDFY1), which serves as an adapter protein in tolllike receptor signaling [68]. Lastly, the gene for dorsal inhibitory axon guidance protein (DRAXIN) was strongly upregulated in L-Palmitoylcarnitine MedChemExpress Epha2-indel722 lenses and that for actin, alpha 2, smooth Rimsulfuron MedChemExpress muscle, aorta (ACTA2) was moderately upregulated in Epha2-null lenses. Whilst ACTA2 serves as a marker for epithelial esenchymal transition in the course of cataract formation [69] and several in the other upregulated genes share cytoskeletal-related or signaling functions, none have but been related with EPHA2 signaling or lens cell differentiation. Amongst the most downregulated genes, two have already been straight implicated in lensspecific cytoskeleton biology. One of the most regularly downregulated gene in Epha2-Q722 (-4-fold), Epha2-indel722 (-100-fold), and Epha2-null (-3-fold) lenses was that for lens glutamine synthase-like or lengsin (LGSN), also called glutamate-ammonia ligase (glutamine synthase) domain containing 1 (GLULD1), a lens-specific protein having a glutamine synthase domain lacking glutamine synthase activity [55]. LGSN is a late marker for lens fiber cell terminal differentiation and has been shown to co-localize with actin and interact using the lens-specific intermediate filament protein, beaded filament structural protein-2 (BFSP2), also referred to as cytoskeletal protein 49 (CP49) or phakinin, suggesting that LGSN represents a recruited enzyme adapted to act as a cytoskeletal component or chaperone in the course of remodeling of your lens cytoskeleton [55,70]. By far the most downregulated gene in Epha2-indel722 mutant lenses (-1000-fold), and to a lesser extent in Epha2-null lenses (-2-fold), was that for chloride intracellular channel 5 (CLIC5). Mutations in the human CLIC5 gene happen to be linked with progressive autosomal recessive, non-syndromic sensorineural hearing impairment with or without the need of vestibular dysfunction and CLIC5 was identified to be abundantly expressed within the fetal inner ear [71,72]. Similarly, in jitterbug (jbg) mice a spontaneous deletion mutation in Clic5 underlies hearing loss with vestibular and renal dysfunction and CLIC5 was localized to the base of hair cell stereocilia where it complexes with radixin, taperin, and myosin VI to stabilize cell membrane ctin cytoskeleton attachments [73]. Lately, CLIC5 been localized to cilia and/or centrosomes inside the lens and Clic5-mutant (jtb) lenses had been found to exhibit defective suture formation [56]. Further, EPHA2 has been shown to regulate Src/cortactin/F-actin complexes during epithelial-to-fiber cell morphogenesis (meridional row and fulcrum formation) at the lens equator [32]. Collectively, these observations point to a functional synergy between EPHA2 and various cytoskeletal proteins with LGSN and CLIC5 delivering promising candidates for future research of EPHA2 signaling within the lens. In conclusion, our information recommend that EPHA2 signaling is expected for lens cell pattern recognition and support a role for EPHA2 in cytoskeleton dynamics through lens cell differentiation.Cells 2021, 10,15 ofSupplementary Components: The following are offered on-line at https://www.mdpi.com/article/ 10.3390/cells10102606/s1. Figure S1. Allele-specific PCR-genotyping of Epha2-mutant mice. (A) PCR ampl.