Additional file 1 of Downregulation of pro-surfactant protein B contributes to the recurrence of early-stage non-small cell lung cancer by activating PGK1-mediated Akt signaling

Additional file 1: Fig. S1 RNA sequencing analysis. a Heatmap showing differentially expressed genes between primary tumors of stage I lung adenocarcinoma patients with postoperative metastatic recurrence and those with no recurrence. RNA sequencing was performed using primary tumors from stage I lung adenocarcinoma patients who experienced metastatic recurrence within 24 months after the operation (n=7) and from patients who experienced no recurrence within 60 months after the operation (n=7). b Genes whose expression was significantly increased in primary tumors from stage I lung adenocarcinoma patients who had not relapsed within 24 months relative to those who had relapsed within 60 months. Fig. S2 Measurement of pro-SFTPB expression in normal lung epithelial cells and NSCLC cell lines by Western blotting. HPA-EpiC, human pulmonary alveolar epithelial cell. Fig. S3 Overexpression of pro-SFTPB inhibited NSCLC progression in animal models. a Vector or pro-SFTPB expression plasmid was transfected into H460 cells. The expression level of pro-SFTPB was measured by Western blot in normal lung epithelial cells and H460 cells that transfected with vector or pro-SFTPB expression plasmid (after 72 hs of transfection). b Subcutaneous xenograft model experiment (n=5/group) showing that overexpression of pro-SFTPB inhibited tumor growth. Subcutaneously inject 1 × 107 H460 cells in 1ml PBS into the back of each nude mouse. Tumors were collected and weighted after 1 month of cell injection. (c) Lung metastasis analysis showing that overexpression of pro-SFTPB expression inhibited tumor metastasis (n=5/group). 1 × 107 H460 cells in 1ml PBS injected into each nude mouse through the tail vein. The lungs were collected after one month of cell injection, and counted the number of tumor on the surface of the lungs. Fig. S4 Measurement of PGK1 expression by Western blotting. H1299 and PC-9 cells were transfected with siRNAs against PGK1. The cells were subjected to Western blotting 72 h after transfection. Table S1. Proteins affected by pro-SFTPB silencing. Table S2. shRNA or siRNA sequences used throughout this study. Table S3 Characteristics of patients with early-stage lung adenocarcinoma (For IHC). Table S4 Characteristics of patients with early-stage lung adenocarcinoma (for serum).