Figure 1. Expression profile of Wnt
ligands and Wnt-related genes in human trabecular meshwork cells. Total
RNA extracted from cells was reverse transcribed and amplified with
specific primers for Wnt-related genes listed in
Table 1.
PCR products were resolved on a 1% agarose gel and visualized by
ethidium bromide staining. Positive PCR products were obtained for
Wnt2b,
Wnt3,
Wnt5a,
Wnt5b,
Dsh1,
Dsh2,
Dsh3,
Fzd1,
Fzd2,
Fzd4,
Fzd5,
Fzd7,
LRP5,
LRP6,
CTNNB1,
GSK3β,
APC,
TCF3,
TCF4,
TCF7,
Dkk1,
Dkk2,
sFRP1,
sFRP2,
and
sFRP3. Negative controls with RNA samples that were not
subjected to reverse transcription were included for all PCR reactions.
No PCR products were seen with any of the negative controls
(representative results are shown for
Wnt2b and
Wnt5a).
When applicable, positive controls, using MCF-7 (shown for
Wnt3
and
Wnt5b) and HEK293 cDNAs, were performed in parallel. All
PCR products were confirmed by sequence analyses, and all the
experiments were performed in at least two different cell lines from
two different donors.