DEV037432 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

• Supplemental Figure S1 -

  Fig. S1. Controls for Dicer and Dgcr8 MOs. (A) Diagram of the GFP-fusion constructs. The positions of the respective MO binding sites are indicated by a bar. AA, amino acids. (B-G) Fluorescence of Xenopus embryos at late gastrula stage injected with Dicer-GFP or Dgcr8-GFP in the presence or absence of Dicer-MO or Dgcr8-MO, as well as uninjected controls. Representative images are shown. Note that both MOs completely abolished GFP expression upon co-injection.

• Supplemental Figure S2 -

  Fig. S2. Time-course of 4A6 staining. (A-C′) Whole-mount immunohistochemistry with 4A6 of uninjected control and Dgcr8-MO-injected Xenopus embryos at stages 40, 42 and 45. Arrowheads indicate the loss of 4A6 staining in the duct that is recovered at later stages of development. Asterisks mark the distal tubular defects. (D) Quantification of 4A6 staining in the pronephric duct from A-C′. Number (N) of the embryos analyzed is indicated above the bars.

• Supplemental Figure S3 -

  Fig. S3. Expression of pronephric marker genes in Dicer morphants. (A-D′) Whole-mount in situ hybridization of uninjected control and Dicer-MO-injected Xenopus embryos at stage 39 with SGLT1-K (A,A′), NKCC2 (B,B′), ClC-K (C,C′) and NCC (D,D′).

• Supplemental Figure S4 -

  Fig. S4. Screening for developmentally regulated and kidney-enriched miRNAs. (A) Microarray analysis comparing miRNA expression in E14.5 and P1 mouse kidneys. (B) RT-PCR expression of candidate miRNAs comparing four adult mouse tissues (kidney, heart, liver, lung) and Xenopus embryos at stages 30, 38 and 45.

• Supplemental Figure S5 -

  Fig. S5. Effects of Xlim1 on 4A6 staining. (A-B′) Whole-mount immunostaining with the 4A6 antibody, comparing the left and right side of Xenopus embryos injected with pCS2-Lhx1* DNA at stage 40. Note the various degrees of change caused by the ectopic expression of Xlim1 (arrows).