Merlin rescue drives Wnt signalling

To investigate the effect of Merlin rescue on cell proliferation in 3D, we grew subcutaneous xenografts from NF2-deficient CH-157MN meningioma cells harboring doxycycline inducible WT-Merlin. Mice were treated with or without doxycycline and Merlin induction was confirmed 24 hours after induction with Western blot. We performed single-cell RNA sequencing on five Merlin-deficient tumors and seven Merlin rescue tumors 3 million CH157-MN meningioma cells hosting doxycycline inducible WT Merlin were injected into the right flank of 4-6wk old NU/NU mice. Doxycycline (20ug/ml) was supplemented in the water of the treatment group at the time of injection. Tumours were dissociated using Collagenase type 7 and 0.25% trypsin. Single cells were processed through a 10X Genomics Chromium Controller and libraries were created using the Chromium Single Cell 3’ Library & Gel Bead Kit v3. Libraries were sequenced on a NovaSeq S4 6000.