library_construction_protocol: 10  anaesthetized larvae were washed twice in ice cold HBSS (Life Technologies GmbH #14175095) and resuspended in 500 l TryplE Mix (1xTrypleE (Life Technologies GmbH #A1217701) in HBSS , 1mM EDTA pH 8 (Molekula #900493) 10 mM 4sU (Merck Chemicals GmbH #T4509) for labelled sample) and incubated at 37°C for 30 min. Enzymes were inactivated by adding 15 µl 10% BSA  (Sigma-Aldrich; #A4503) before spinning down and resuspending the cells in TRIzol RNA was extracted using TRIzol-Chloroform-Isopropanol according to the manufacturer's instructions. Iodiacetamide Treatment was carried out on extracted RNA in 50% DMSO as described in Herzog et. Al 2017 (https://doi.org/10.1038/nmeth.4435). libraries for RNA-seq were prepared following a CELseq protocol as published in Holler & Junker 2019 (https://doi.org/10.1007/978-1-4939-9009-2_9). Sequencing adapters were switched – Insert is in read 1!