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. 2023 May 23;3(6):100488. doi: 10.1016/j.crmeth.2023.100488

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Optimization of 2D BMEx overlay method

(A) Experimental scheme depicting the different conditions tested in (B) and (C).

(B) FACS plots depicting the percentage of double EPCAM+ITGA6+ cells at day 5 in line F99 to test different BMP4 concentrations.

(C) Immunofluorescence for POU5F1 and SOX17 at day 5 in line F99 to test different BMP4 concentrations. Dashed box is magnified (bottom), showing separated channels. Scale bars: 50 μm.

(D) Experimental scheme depicting different conditions tested (left) and the associated FACS plots (right) depicting the percentage of double EPCAM+ITGA6+ cells at day 5 in line F99.

(E) Representative FACS plots showing percentages of double EPCAM+ITGA6+ cells at day 5 generated using Geltrex, Cultrex, and Matrigel overlay in line M54.

(F) Bar graph showing mean percentages of double EPCAM+ITGA6+ cells at day 5 generated using different brands and batches of BMEx, analyzed by FACS in line M54; error bars represent mean ± SD.

See also Figure S1.