Xenbase Image ID: 129058

Figure 3. Oxysterols and 22-NHC bind the extracellular cysteine-rich domain (CRD) of vertebrate Smo(a) Secreted HA-tagged mSmoCRD and detergent extracts of 293T cells expressing mSmo-Cherry were tested for binding to 22-NHC beads, in the presence of free 22-NHC. MSmoCRD binds 22-NHC beads, similar to mSmo.(b) MSmoCRD binding to 22-NHC beads was assayed as in (a), in the presence of 7-OHC, 20-OHC or 20-OHC-Pent. Only the active sterols 20-OHC and 20-OHC-Pent compete mSmoCRD binding to 22-NHC beads, while the inactive 7-OHC does not.(c) As in (b) but in the presence of the diastereomers 20(S)-OHC-Pent and 20(R)-OHC-Pent. Only the active S diastereomer competes mSmoCRD binding to 22-NHC beads.(d) MSmoCRD and mSmo binding to 20-OHC beads was assayed as in (c). Both mSmoCRD and mSmo bind 20-OHC beads and are competed by 20(S)-OHC-Pent, but not 20(R)-OHC-Pent. The binding affinities of mSmoCRD and mSmo to 20-OHC beads are similar.The full immunoblots for this figure are shown in Supplementary figure 11. Image published in: Nedelcu D et al. (2013) Image downloaded from an Open Access article in PubMed Central. Image reproduced on Xenbase with permission of the publisher and the copyright holder. Larger Image Printer Friendly View

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