Fig 1: Dose-dependent myelin wrapping of AAs in response to compounds. (A) 3-point series of concentration (taken from 9-point dose response data) illustrate compounds dose-dependent effect on myelin wrapping. (B–D) Top panel: examples of sigmoidal dose–response curves fitted to the wrapping index data for the 9-point concentration series, for, tasin-1, clemastine and T3. The curves for all the tested compounds are shown in Supplementary Fig. S1; The sigmoidal fit excludes concentrations above which the drug induced cytotoxicity. Reported are average values over all fields of view (3 replicates with combined 27 fields of view). Error bars are standard error of the mean (SEM); Middle panel: examples of MBP staining of myelin wrapping around artificial axons (shown is one field of view at a selected z-plane) corresponding the maximum efficacy dose for tasin-1 (111 nM), clemastine (3 µM) and T3 (111 nM); Bottom panel: processed images from middle panel showing wrapped myelin membrane around artificial axons. Scale bars: main image: 200 μm, insert: 100 μm.
Fig 2: (A) The EC50 values (left scale for the compound subset left from the middle line and right scale for the compound subset right from the middle line) and (B) the efficacy values relative to T3, obtained from 9-point dose–response data for wrapping index in 3D myelin wrapping assay (red columns) and for MBP area in 2D differentiation assay (blue columns). The compounds are organized according to the values for 3D wrapping assay, for easier visualization of the different outcomes from the myelin wrapping and differentiation assays. Reported are average values over all fields of view (3 replicates with combined 27 fields of view). #EC50 for fasudil in 3D AA assay was not determined due to the lack of plateau of dose response curve at high dose range; ##EC50 for bazedoxifene in 2D differentiation assay: 0.1 nM. In (B) error bars are standard error of the mean (SEM), numbers denote rank, horizontal lines denote 2 × standard deviation for T3 in the respective 3D and 2D assays; In (A) EC50 values are obtained from one experimental screen with 3 replicates resulting in one dose–response curve, therefore no SEM is available. *p-value < 0.05 between 3 and 2D conditions for each compound.
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