4 files

Supplemental Material for Deng et al.

posted on 12.09.2019, 14:44 by Yuting Deng, Katherine Leisan Luo, Daniel D. Shaye, Iva Greenwald

Supplemental Table S1. Strains used in this work.

Supplemental Table S2. List of conserved kinases screened to find new negative regulators of LIN-12/Notch. Validated positive hits are highlighted in blue. The WormBase ID (WBgene), common name and locus ID for the C. elegans kinases screened are shown in the first three columns. The fourth column shows either (i) feeding-RNAi clones obtained from the commercially available “Ahringer Library” (Kamath and Ahringer 2003), (ii) clones derived from the “Vidal Library” (Rual et al. 2004), and (iii) clones generated by us to fix those that were not correct in the Ahringer Library (against daf-2, gck-1 and T01G5.1), and supplemental clones that did targeted all predicted isoforms of certain genes (atl-1, pkc-1) or did not exist in either the Ahringer or Vidal libraries. The bottom of the table shows clones that targeted five kinase-encoding genes that are now considered pseudogenes (www.wormbase.org, ver. WS272).

Supplemental Table S3. Primers used to make RNAi clones. We used these primers to PCR amplify fragments from genomic DNA that encompass exonic regions to target kinases. Most of the primers, and resulting PCR products, described here were made to generate clones against kinases that were not represented in either of the available “feeding” RNAi libraries (see main text). Three of the PCR products were used to correct erroneous clones (daf-2, gck-1 and T01G5.1), or to create new clones that would target kinase isoforms that were missed by previously existing clones (atl-1 and pkc-1). The bottom portion of the table shows primers used to build second, or third, “non-overlapping” RNAi clones to confirm results from the initial screen for those genes where null alleles were unavailable or caused lethality (see Fig. 3, main text).

Supplemental Figure S1. Additional biological replicate testing enhancement of lin-12(n302) as compared to lin-12(+).


Article title

A Screen of the Conserved Kinome for Negative Regulators of LIN-12 Negative Regulatory Region ("NRR")-Missense Activity in Caenorhabditis elegans

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