Fig. S6

Western blot of 6 d wild-type (nonTg WT), Crb2a^IntraWT, Crb2a^IntraΔFBD, Crb2a^IntraΔPBD, Crb2a^{IntraΔFBDΔPBD}, Crb2a^FL, Crb2a^Extra_TM, Crb2a^Extra_Secr transgenics probed with anti-HA antibodies. Western blotting was performed as previously described [8] and probed with anti-HA (clone 16B12) and HRP-conjugated goat anti-mouse. The predicted molecular weight of Crb2a^IntraWT protein is 11 kD (retaining the signal peptide) but on western blot the major Crb2a^IntraWT protein is ∼30kD, suggesting that it may be post-translationally modified or forms homoligomeres. Despite trying multiple gel and transfer conditions we were unable to detect Crb2a^IntraΔFBD, Crb2a^IntraΔPBD, Crb2a^{IntraΔFBDΔPBD} proteins, which by immunohistochemistry are expressed at similar levels as Crb2a^IntraWT. It is possible that Crb2a^IntraΔFBD, which is be predicted to be about the same molecular weight as Crb2a^IntraWT, is not post-translationally modified or does not dimerize and, thus, is too small, like Crb2a^IntraΔPBD and Crb2a^{IntraΔFBDΔPBD} with predicted molecular weights ∼8.8kD (with signal peptide) to be captured by Western blot analysis.