Journal: Histochemistry and Cell Biology
Article Title: Spatio-temporal distribution of tubulin-binding cofactors and posttranslational modifications of tubulin in the cochlea of mice
doi: 10.1007/s00418-020-01905-6
Figure Lengend Snippet: Distribution of TBC proteins in the organ of Corti at P1 and P7. a – e age P1; a TBCA expression is found in the inner area of Kölliker’s organ, the inner hair cell and a tectal cell. b TBCB is expressed in the cells of the Kölliker’s organ. c TBCC labelling is detected in the basal half of the two pillar cells. d Similar TBCD is expressed in the pillar cells. e TBCE staining is found in the basal cells of Kölliker’s organ and the basal half of the inner phalangeal cell. Diffuse staining is visible around the nucleus and below the apical surface of the inner hair cells. The three outer hair cells are also marked. f – j age P7: f TBCA is detected in the inner and outer pillar cells, phalangeal extensions of the three Deiters’ cells, tectal cells and Hensen’s cells. g Similar to TBCA, TBCB is detected in the inner and outer pillar cells, the three Deiters’ cells, the tectal cells and Hensen’s cells. h The expression of TBCC is found in the supporting cells (inner and outer pillar cells, the phalangeal extensions of the three Deiters’ cells, the tectal cells and Hensen’s cells). i TBCD staining is found in the basal parts of the inner pillar cell. j In addition to the inner pillar cells, TBCE is also detected in the and the basal half of the inner phalangeal cell phalangeal extensions of Deiters’ cells. ( TM tectorial membrane with unspecific staining) (Scale bar = 25 µm)
Article Snippet: For immunofluorescence analysis, sections were post-fixed in 4% PFA for 10 min, rinsed in 0.05 M phosphate-buffered saline three times and blocked in 10% bovine serum albumin (BSA) in 0.1% Triton X-100 for 30 min. Primary antibodies were incubated in 1% BSA in 0.1% Triton X-100 overnight at 4 °C at the following concentrations: rabbit polyclonal against TBCA (1:50), TBCB (1:100), TBCC (1:200), TBCD (1:100) and TBCE (1:100, all Proteintech), mouse monoclonal against tyrosinated tubulin (1:500, Millipore Merck), rabbit polyclonal against detyrosinated tubulin (1:500, Millipore Merck), mouse monoclonal against acetylated tubulin (1:500, Sigma-Aldrich) or mouse monoclonal against polyglutamylated tubulin (1:500, AdipoGen).
Techniques: Expressing, Staining, Membrane