I have been trying to get Ciliobrevin D to inhibit dynein motor activity in primary neuronal (cortical and dorsal root ganglion) cultures. I am tracking axonal transport of lysosome as a proxy for dynein activity (Ref: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4436090/). I dissolved Cil D in DMSO at 50 mM and stored stocks aliquotted at - 20 less than 2 months ago. I used it at a final concentration of 25 uM or 50 uM and treat cells for ~ 45 min before tracking lysosome transport. But there is no inhibition of transport in both cortical and DRG cultures. The imaging was done at 37C with 5% CO2. I also tried Ciliobrevin A at 60 uM under the same conditions but had no luck with that either. Please share if you have experienced this problem before or have any suggestions or a different drug that has worked for you. Thanks!