Defining signals that promote human alveolar type I differentiation-Supplementary Figures

<p><strong>Supplemental Figure 1. Human serum robustly promotes alveolar type I differentiation in part through BMP signaling pathway</strong></p><ol><li>Schematic for generating ATI spheroids using human serum.</li><li>The inhibition of <i>AGER</i> gene expression by Noggin under ADM containing human serum (*<i>P</i> &lt; 0.05 and **<i>P</i> &lt; 0.01).</li></ol><p>C-E. Immunostaining for SFTPC and AGER for ATII spheroids treated with ADM without human serum, ADM and ADM plus Noggin. Representative of&nbsp;three independent experiments. Scale bar: 50µm.</p><ol><li>Quantification of SFTPC and AGER for ATII spheroids treated with ADM without human serum, ADM and ADM plus Noggin. Representative of&nbsp;three independent experiments.</li></ol><p><strong>Supplemental Figure 2. BMP4 promoted </strong><i><strong>HOPX</strong></i><strong> transcript expression in ATII spheroids in a BMP concentration-dependent manner. Representative of&nbsp;three independent experiments</strong></p><p><strong>Supplemental Figure 3. CDKN1A and CLDN4 expression in ATII spheroids cultured under hATIIM, hATIIM + BMP4 and hATIDM.</strong></p><ol><li>Schematic for CDKN1A and CLDN4 expression using ATII spheroids under hATIIM, hATIIM + BMP and hATIDM.</li><li>The <i>CDKN1A</i> and <i>CLDN4</i> transcript expression under hATIIM, hATIIM + BMP and hATIDM in three biological samples and triplicate technical repeat.</li></ol><p>C-E. Immunostaining for CDKN1A, SFTPC and AGER for ATII spheroids cultured with hATIIM, hATIIM + BMP and hATIDM. Representative of&nbsp;four independent experiments. Scale bar: 50µm.</p><p>F. Quantification of CDKN1A for ATII spheroids treated with hATIIM, hATIIM + BMP and hATIDM. Representative of&nbsp;four independent biological samples.</p><p><strong>Supplemental Figure 4. The ATI differentiation efficiency comparison of ATII spheroids and DLOs using hATIDM and sc-RNA Sequencing</strong></p><ol><li>The relative expression of <i>SFTPC</i>, <i>AGER</i> and <i>HOPX</i> using sc-RNA sequencing from http://ipfcellatlas.com/</li></ol><p>&nbsp; &nbsp; &nbsp; &nbsp;B-D. The relative expression of <i>SFTPC</i>, <i>AGER</i> and <i>HOPX</i>from ATII spheroids and DLOs using hATIIM and hATIDM,&nbsp;</p>