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. Author manuscript; available in PMC: 2014 Dec 9.
Published in final edited form as: Nat Commun. 2014 Jun 9;5:4088. doi: 10.1038/ncomms5088

Figure 5. Tcf3 promotes cell migration non-cell autonomously and independently of its role as a cofactor to β-catenin.

Figure 5

(a) Real Time PCR analysis of level of expression of transduced tet-inducible Tcf3 and its mutant versions, Tcf3ΔN and Tcf3ΔG, in response to Dox. Keratinocytes were transduced with GFP-tagged, tet-inducible lentivectors carrying myc-tagged Tcf3 or its mutant versions. Transduced cells were isolated by fluorescent cell sorting and were treated with Dox or vehicle control for 24 hours prior to being harvested for mRNA analysis. Mean expression values from biological duplicates were calculated. Data are mean ± s.d (Student’s t-test).

(b) Images of transduced keratinocytes expressing tet-inducible Tcf3, Tcf3ΔN, Tcf3ΔG 16 hrs after the initiation of migration assay with or without Dox. Black bar denotes 200μm.

(c) Graph quantifying the area migrated by transduced cells treated with Dox relative to area migrated by cells treated with vehicle control. For each sample, over 30 non-overlapping fields were measured at each timepoint; and each experiment was repeated twice. Data are mean ± s.d. **p<0.001 (Student’s t-test).

(d) Tcf3-induced cells secrete factors that promote cell migration. Keratinocytes from tetinducible Tcf3 (K14-rtTA;TRE-mycTcf3) and control (K14-rtTA) mice were cultured in the presence or absence of Dox for 48 hours and their conditioned media were collected. Wild type keratinocytes were subjected to a migration assay in the presence of specified conditioned media. Images were taken 38hrs after the initiation of migration assay. Bar denotes 200μm.

(e) Conditioned media from Tcf3-induced cells (K14-rtTA;TRE-mycTcf3 +Dox) promotes migration of wild-type cells. Graph quantifying the migrated area of wild-type cells that were incubated with Dox treated conditioned media during the migration assay relative to the area migrated by cells treated with vehicle treated conditioned media. For each sample, over 30 non-overlapping fields were measured at each timepoint; and each experiment was repeated three times. Data are mean ± s.d. **p<0.001 (Student’s t-test)..