Dapta

The Animal Care and Use Committee at Xinhua Hospital, Shanghai, China approved all mouse and rat protocols. We used a previously established animal model of BPD^{10 (link),11 (link)}. Briefly, at 16.5 days of gestation (term: 22 days), pregnant rats were prepared to receive intra-amniotic injection and were randomly assigned to the saline control, LPS, LPS with batyl alcohol (BTA), LPS with D-ala-peptide T-amide (DAPTA), LPS with anti-IL-1β antibody (anti-IL-1β), LPS with GSK872 (RIP3 kinase inhibitor) or IL-1β group. The saline control group received 5 μL of normal saline per amniotic sac, the LPS group while the LPS, LPS with BTA, LPS with DAPTA, LPS with anti IL-1β, LPS with GSK872 and IL-1β groups received 1 μg of LPS (Escherichia coli 055: B5; Sigma-Aldrich, St. Louis, MO, USA), 1 μg of LPS and 0.4 μg of BTA (Bachem, Bubendorf, Switzerland), 1 μg of LPS and 1 μg of DAPTA (Tocris Bioscience, Bristol, UK), 1 μg of LPS and 0.05 μg of anti-IL-1β (PeproTech, Rocky Hill, NJ, USA), 2 μg of GSK872 (Selleckchem,Houston, TX, USA), and 0.5 μg of IL-1β (PeproTech), respectively, diluted in 5 μL normal saline per amniotic sac. The day the pups were born was designated as postnatal day 0 (P0). Both male and female rats were used, and animals were killed on P1, P3, and P7 to examine the time course changes for all measurements.

DAPTA was purchased from Tocris Bioscience (Bristol, UK). Fluorescein isothiocyanate, PE/Cyanine7, Allophycocyanin, Alexa Fluor^® 488, PE/Dazzle™ 594, Alexa Fluor^® 647, and phycoerythrin-labeled CD40, NF-κB p65, IκBα, Notch1, Notch3, GM-CSF, iNOS, RORγT, MCP-1, TNF-α, lysis, and permeabilization solution, and fixation buffers were purchased from BD Biosciences and BioLegend (San Diego, CA, USA). SYBR Green and cDNA kits were purchased from Applied Biosystems (Foster City, CA, USA). Primers were purchased from GenScript (Piscataway, NJ, USA). TRizol was purchased from Invitrogen (Carlsbad, CA, USA).