(A) Validated screening hits from Li et al. 2022 with annotations of subsequently-established inhibitory activity for EBP, CYP51, or sterol 14-reductase. (B), (E), (F) GC-MS based quantification of zymostenol and zymosterol levels in GBM528 cells treated for 24 h with the indicated screening hits at 10 μM. (C) GC-MS based quantification of lanosterol levels in GBM528 cells treated for 24 h with clotrimazole at 10μM. (D), (G) GC-MS based quantification of 14-dehydrozymostenol levels in GBM528 cells treated for 24 h with the indicated molecules at 10 μM. n=2 wells per condition. (H) Percentage of MBP+ oligodendrocytes generated from OPCs following treatment with bioactive small molecules in dose response as shown. n=4 wells per condition, except DMSO and amorolfine, n=8 wells with >1,000 cells analyzed per well.