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Figure 6. (A) Schematic of the induction protocol used to generate hNCCs in which hESCs were exposed to CHIR 99021 for 2 days in serum-free medium prior to withdrawal of this agent and continued culture for 3 additional days. (B) Immunofluorescent antibody staining for NCC markers PAX7 and SOX10 at day 5 in EMC1 siRNA-treated cells as compared with control siRNA-treated cells performed in 2 separate biological replicates. Scale bar: 100 μm. (C) qPCR analysis at day 2 and day 5 showed a decrease in transcripts of EMC1 performed in 2 separate biological replicates. (D) qPCR analysis at day 5 performed in 2 separate biological replicates showed decreased levels of several NCC marker transcripts (PAX7, SNAI2, SOX9, FOXD3, SOX10), although other markers appeared to be unaffected (PAX3). (E) Quantified percentage of cells expressing each PAX7 and SOX10 protein performed in 2 separate biological replicates. Image published in: Marquez J et al. (2020) Copyright © 2020. This image is reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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