Fig. S3

Lypd6 is required for Wnt/β-catenin signaling in Wnt-receiving cells.
related to Figure 4.

(A) Mouse lypd6 transcripts are knocked down to about 27% in murine L cells transfected with lypd6 siRNA as determined by qRT-PCR at 48 hours after transfection.
(B) Human lypd6 transcripts are knocked down to about 23% in HEK293T cells transfected with lypd6 siRNA as determined by qRT-PCR at 48 hours after transfection.
(C) Human lypd6 transcripts are knocked down to about 5% in HEK293T cells transfected with lypd6 esiRNA as determined by qRT-PCR at 48 hours after transfection.
(D) siRNA mediated knockdown of human lypd6 in HEK293T cells reduces induction of pBAR by Wnt3a conditioned medium (CM). pBAR activity is shown relative to that in cells transfected with negative control siRNA and treated with control CM.
(E) esiRNA mediated knockdown of human lypd6 in HEK293T cells reduces induction of pBAR by Wnt3a CM.
(F) siRNA mediated knockdown of human lypd6 in RKO cells reduces induction of pBAR by Wnt3a CM. (A-F) Error bars, s.e.m.
(G) otx2 expression (blue) in zebrafish host embryos transplanted with wnt8 expressing cells, marked by fluorescein-tagged dextran detected in brown at 100% epiboly. D: donor, H: host, anterior views.
Upper row: Control non-transplanted embryo injected with control MO and control RNA (n=15/15).
Middle row: wnt8 expressing cells, transplanted into a wild type (uninjected) embryo at dome stage, can repress otx2 (blue) no matter whether the donor cells have been co-injected with lypd6 MO (n=8/11) or control MO (n=12/13).
Bottom row: wnt8 expressing cells inhibit otx2 expression in control MO injected hosts (n=8/9), but much less so if the host was injected with lypd6 MO (n=10/13).