DEV003194 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. RT-PCR was performed on cDNA synthesised from RNA extracted from mammary glands collected at day 5, 10 and 15 of gestation, and at day 0, 5 and 10 of lactation from WT and KO mice. Specific primers for Il4ra1, Il13ra1, Gata3, Gapdh and cyclophilin A were designed using Quick-prime program and the sequences were checked for specific binding using blast-n search engine.

• Supplemental Figure S2 -

  Fig. S2. Normal pubertal development of the mammary gland in Stat6^-/- mice. Inguinal mammary glands were collected at 5 weeks of age for whole-mount analysis. LN, lymph node; TEB, terminal end buds migrating into the fat pad.

• Supplemental Figure S3 -

  Fig. S3. Progesterone receptor, prolactin receptor, and insulin growth factor I and II expression is not affected in Stat6 KO mammary glands. RT-PCR was performed on cDNA synthesised from RNA extracted from mammary glands collected at day 5, 10 and 15 of gestation and at day 0, 5 and 10 of lactation from WT and KO mice. Specific primers were designed using Quick-prime program and the sequences was checked for specific binding using blast-n search engine.

• Supplemental Figure S4 -

  Fig. S4. The kinetics of IL-4 and IL-13 expression during gestation and lactation. Error bars represent s.d. of three experiments

• Supplemental Figure S5 -

  Fig. S5. Expression of IL-4/IL-13/Stat6 signalling components in KIM-2 cells. (A) Morphology of KIM-2 cells at confluency and at day 2 of differentiation. (B) RT-PCR showing that KIM-2 cells express IL-4Ra, IL-13Ra1 and Gata3 across a differentiation time course. (C) Immunoblots for pStat6, Stat6, Gata3, c-Maf, β-casein and α-tubulin.

• Supplemental Figure S6 -

  Fig. S6. H&E staining of mammary gland sections from WT and Stat6^-/- mice at day 0, 5 and 10 of lactation.