Figure S1. Pilot Study for the Functional Screening and the Specificity Test for XFDL156-MO (A-D) Poly A+ RNAs were prepared from animal cap cells of stage 8.5 and stage 11.5. Control (10 ng/cell; A,B), stage 8.5 (10 ng/cell; C) and stage 11.5 (10 ng/cell; D) mRNAs were injected at the 4-cell stage. Animal caps were prepared at stage 8.5, cultured in the presence of 5 ng/ml of Activin (B-D), harvested at stage 11 (equivalent) and analyzed for Xbra expression by in situ hybridization. (E-J) Immunohistochemistry using a preimmune antiserum (E) or XFDL156 antiserum (H) in stage-10.5 animal cap cells. DAPI staining is shown in green. Merged views are shown in (G) and (J). (K) XFDL156-MO and XFDL141-MO (for the short variant) specifically inhibited protein synthesis from each transcript. Synthetic XFDL156 and XFDL141 mRNAs were prepared in vitro from pCS2-Venus, pCS2-5TR-XFDL156 (lanes 2-5) and pCS2-5TR-XFDL141 (lanes 6-9) without (lanes 2 and 6) or with 0.25 μg/μl (lane 3) or 1 μg/μl (lane 4) of XFDL156-MO, 1 μg/μl of 5-mismatched XFDL156-MO (lane 5), 0.25 μg/μl (lane 7) or 1 μg/μl (lane 8) of XFDL141-MO or of 5-mismatched XFDL141-MO (lane 9). The translated proteins were labeled with [35S]-Methionine, resolved by SDS-PAGE and detected by autoradiography.
Image published in: Sasai N et al. (2008)
Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V.
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