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. 2022 May 17;39(7):110811. doi: 10.1016/j.celrep.2022.110811

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© 2022 The Authors

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

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SHH signaling is up-regulated in INPP5E^D477N/D477N organoids

(A and B) Examples of differentially expressed dorsal and ventral (A) and signaling factor genes (B) with indicated log2Fold changes in D24 organoids.

(C, D, F, and G) In situ hybridization to detect PTCH1 and GLI1 expression in D39 control and INPP5E mutant organoids.

(E and H) Representative example of qRT-PCR analyses showing PTCH1 (E) and GLI1 (H) mRNA expression relative to ATP5 at D24.

(I) GLI3 western blot on D24 organoid tissue revealed the GLI3 full-length (FL) and repressor (R) forms.

(J–L) Quantification of GLI3 western blot. GLI3R levels are decreased in INPP5E^D477N/D477N organoids, while GLI3FL levels and the GLI3R/GLI3FL ratio (L) are not affected.

All statistical data are presented as means ± SD; unpaired t tests with Welch’s correction (n = 3 for control and n = 2 for mutant; E and H); unpaired t tests (J and K); Mann-Whitney test (L) with n = 3 for control and n = 4 for mutant; ^∗p < 0.05. Scale bar, 100 μm (C). See also Figures S4 and S5.