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Figure 2 | BMC Developmental Biology

Figure 2

From: Delta activity independent of its activity as a ligand of Notch

Figure 2

Dl down-regulates Daughterless protein expression, and N up-regulates E(spl)C m3 gene expression, in response to Scabrous. A. Northern blots of total RNA from the indicated cell mixtures extracted at 0 or 45 minutes after treatment with medium containing Sca (+) or not (-). Gene probes used are shown on the right. m3 = E(spl)C m3 and rp 49 = a ribosomal protein gene used to show the levels of total RNA in the lanes in all northern blots. Sca = conditioned medium prepared from the S2-Sca stable cell line in all experiments here onwards. The control medium used along side Sca medium (-) was prepared from heat shocked S2 cells. Experiments were repeated two times. For unknown reasons, the medium collected from heat shocked S2 cells (used in lanes 1, 3, and 5) produced higher background levels of E(spl)C m3 RNA in S2-N cells (lane 1). B. Western blots showing the levels of Da and Dl in different Dl cell lines. S2-Dl, S2-Dl(1) and S2-Dl(2) are independently established hsDl cell lines. Ui = un-induced (i.e., not heat shocked). Hsp70 = heat shock 70 protein used to show the levels of proteins in the lanes of all western blots. Dl and DlΔI were detected with αDlEC. Da signals here (and the indicated signals elsewhere) were quantified relative to Hsp70 (western blots), rp49 (northern blots), or other indicated molecules, using the NIH Image 1.63 program. These experiments were repeated more than ten times. C. Western blots showing Da levels in the indicated cell mixtures, with (+) or without (-) Sca. These experiments were repeated five times.

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