Fig. 8
IFITM1 functions as a regulator of metastasis and polarization of M2d-TAM in p-FoxM1-induced metastasis. a, b IFITM1 was depleted in A549S25E cells using human IFITM1 shRNA for 48 h. a qRT-PCR was performed for IFITM1, FOXM1, CDH2, PLK1, TGFB1, IL6, and VEGFA. b Immunoblot analyses were performed using anti-IFITM1, anti-RFP, anti-E-cadherin, anti-p-Smad2, anti-Smad2/3, anti-SNAI1, anti-SNAI2, and anti-GAPDH antibodies. c, d 5 ng/mL TGF-β was applied to A549 cells depleting IFITM1 with shRNA. c Immunoblotting was performed using anti-IFITM1, anti-FoxM1, anti-E-cadherin, anti-N-cadherin, anti-p-Smad2, anti-Smad2/3, anti-SNAI2, anti-vimentin, and anti-GAPDH antibodies. d qRT-PCR was performed for IFITM1, FOXM1, CDH2, PLK1, TGFB1, IL6, and VEGFA. e IFITM1 was depleted in A549S25E cells using human IFITM1 shRNA for 48 h. Cells were subjected to a Transwell migration assay. Three days after seeding, the cells on the bottom surface were stained with 0.05% crystal violet dye. Images of the Transwell cell migration assay were collected and analyzed with an Odyssey infrared imaging system and plotted. f An invasion assay was performed using A549 cells expressing wild-type or mutants of FoxM1. Seven days after seeding, the cells that invaded the bottom surface were stained with 0.05% crystal violet dye, and the relative absorbance was plotted (n = 3). g THP-1 cells were co-cultured with A549S25E cells depleting IFITM1 for 48 h. qRT-PCR was performed for CD163, CD206, and VEGFA. h The overall survival (OS) of all LUAD patients (n = 719) (h, upper) and stage 3 LUAD patients (n = 24) (h, lower) was analyzed according to IFITM1 expression level using KM PLOTTER. High (Hi) and low (Lo) were generated based on the expression at the median cut-off. i The OS of all LUAD patients (n = 703) (i, left) and stage 3–4 LUAD patients (n = 137) (i, right) was analyzed according to IFITM1, FOXM1, and PLK1 expression levels using KM PLOTTER. High (Hi) and low (Lo) were generated based on the expression at the median cut-off. Data are presented as mean ± SD of three independent experiments (significantly different from the experimental control). *p < 0.05; ***p < 0.001; (n = 3)