Figure 5. TPPP KO Oligodendrocytes Have Microtubule Organization Defects.
(A) Micrographs of DIV4 oligodendrocytes cultured from WT or TPPP KO mice and immunostained against tubulin.
(B) Sample traces of tubulin-stained oligodendrocytes used for Sholl analysis.
(C) Sholl analysis of microtubule processes in WT versus TPPP KO oligodendrocytes.
(D) Width of primary processes. * p = 0.014. n = 13–20 cells, 4 mice per genotype.
(E) Kymographs from WT versus TPPP KO oligodendrocytes expressing mCherry-EB3 (DIV4).
(F) Percent of microtubule plus-ends labeled with mCherry-EB3 that are growing away from the cell body in DIV4 oligodendrocytes. *** p = 5 × 10−9.
(G) Microtubule growth speeds of mCherry-EB3 in DIV4 oligodendrocytes. p = 0.52.
(H) Microtubule growth length of mCherry-EB3 in DIV4 oligodendrocytes. p = 0.11.
(I) Flux of anterograde mCherry-EB3 growing microtubules in distal processes (>15 μm from the cell body). * p = 0.05. n = 9–10 cells, 3 mice per genotype.
(J) Model for local microtubule polarity and branching regulation at Golgi outposts by TPPP.
(K) Number of Golgi outposts per oligodendrocyte. n = 11–17 cells, 3 mice per genotype. * p = 0.05.
(L) Micrographs of TPPP KO oligodendrocyte immunostained against GM130 and tubulin.
(M) Micrographs of smFISH against mouse Mbp mRNA.
(N) Number of Mbp mRNA aggregates with size >2 μm. ** p = 0.001.
(O) Size of Mbp mRNA aggregates. ** p = 0.004. n = 18–29 cells, 3 mice per genotype.