th aves labs cat tyh 0020  (AvesLabs)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: TMEM63A, associated with hypomyelinating leukodystrophies, is an evolutionarily conserved regulator of myelination

doi: 10.1073/pnas.2507354122

Figure Lengend Snippet: Delayed myelination in Tmem63a -null mice recapitulates the HLD19 phenotype. ( A ) Representative micrographs of 50 µm-thick cortical sections immunostained against MBP (magenta) from Tmem63a WT/WT (gray labels) and Tmem63a KO/KO (rose labels) mice at indicated ages. (Scale bar, 100 µm.) ( B ) MBP coverage in cortical sections, as MBP+ area by total area, for Tmem63a WT/WT and Tmem63a KO/KO mice. Inset region of MBP measurement in the cortex (n = 4 to 6 animals per genotype and age, Brown–Forsythe and Welch ANOVA for multiple comparisons). ( C ) Internode myelin sheath lengths as MBP+ distance between CASPR labeling ( Inset ), for Tmem63a WT/WT and Tmem63a KO/KO mice. (n = 4 to 6 animals per genotype and age, One-way ANOVA for multiple comparisons). (Scale bar, 5 µm.) ( D ) Representative immunoblots of fractions isolated from brain tissue of P30 – P90 Tmem63a WT/WT and Tmem63a KO/KO mice. Blots probed for lysosomal protein LAMP2, myelin-associated protein MBP, and TMEM63A. Lanes: 1) postnuclear supernatant [PNS], 2) cytosol, 3) total cellular membranes [inclusive of ER/PM and lysosomes], 4) ER/PM. Quantification of LAMP2 ( E ), MBP ( F ), TMEM63A ( G ) immunoblot signal density normalized to Ponceau S signal density measured between 100 to 150 kDa (n = 8, 2-way ANOVA and Tukey’s multiple comparison testing).

Article Snippet: Labeling was performed overnight at 4 °C with primary antibodies diluted in blocking solution: mouse IgG2a anti-CASPR 1:250 (Millipore Sigma, cat# MABN69), chicken anti-MBP 1:200 (Aves Labs, cat# MBP).

Techniques: Labeling, Western Blot, Isolation, Comparison