PHLOWER - Single cell trajectory analysis using Decomposition of the Hodge Laplacian [Xenium]

We performed multiplex siRNA knockdown experiments of the identified neuronal lineage defining TFs PAX3, RFX4 and ZIC2 in kidney organoids in iPSC derived kidney organoids during the differentiation process from day 19 onwards. Kidney organoids (at least N=3 per condition per experiment, 2 independent experiments) were transfected using ON-TARGETplus™ SMARTpool siRNA’s ZIC2 (Catnr L-017505-00-0005), RFX4 (catnr L-013577-00-0005) and PAX3 (Catnr L-012399-00-0005, work concentration 25nM, Horizon Discovery, Cambridge, UK) and DharmaFECT Transfection reagent 1 (0.5% v/v) from day 7+5 (intermediate mesoderm stage) onwards until the end of the protocol until day 7+18. Organoids were refreshed every other day.