Wandering third instar larvae expressing Disc\RFP^{dsRNA.mCherry.UAS.VALIUM20} under the control of Scer\GAL4¹¹⁵¹ were harvested between 110 and 135 h after egg-laying. The notum and hinge regions of wing discs were dissected and cells were dissociated with collagenase and trypsin in Rinaldini solution.

Single-cell RNA-seq libraries were prepared using the DROP-seq method (Macosko et al., 2015).

RNA-seq libraries were sequenced on the Illumina NextSeq platform.