Figure 2.

Immobilized WNT3A Surfaces Can Be Used to Induce WNT Signaling in a Variety of Stem Cell Cultures

(A) Brightfield (BF) images of Comma Dβ-Geo cells on WNT3A immobilized surfaces with and without DTT treatment, along with corresponding mCherry (SV-40) and GFP (7xTCF) expression.

(B) Percent GFP⁺ cells determined using automated protocol generated in Volocity software; based on finding overlap of mCherry and GFP objects. n = 4 independent experiments, mean ± SD; statistical significance determined with one-way ANOVA, ^∗p < 0.05, ^∗∗p < 0.01.

(C) Brightfield images of 7TCF-eGFP mESC colonies and corresponding GFP expression.

(D) Percent of GFP expressing cell clusters on WNT3A or DTT-treated surfaces. Number of colonies determined manually while GFP⁺ objects found using automated protocol generated in Volocity software. n = 4 independent experiments, mean ± SD; statistical significance determined by unpaired t test assuming equal SD; ^∗∗∗∗p < 0.0001.

(E) Brightfield images of hMSCs expressing 7xTCF-GFP/SV40-mCherry and the corresponding mCherry and GFP expression. Representative images from cells seeded on BSA control, BSA (supplemented with soluble WNT3A), or immobilized WNT3A.

(F) The percent of GFP⁺ cells for each condition; determined using automated protocol generated in Volocity software; based on finding overlap of mCherry and GFP objects. n = 3 independent experiments, mean ± SD; statistical significance determined by two-way ANOVA; ^∗∗p < 0.01, ^∗∗∗p < 0.001; ns, not significant.

The scale bar represents 50 μM.