Fig 1: Effect of si‐PTK7 on NSCLC cell growth, invasion, and macrophage M2 polarization. A549 and H1299 cells were transfected with si‐NC/si‐PTK7. (a) PTK7 protein expression was tested by WB. CCK8 assay (B), EdU assay (c), flow cytometry (d), and transwell assay (e,f) were used to measure cell proliferation, apoptosis, and invasion. (g) CD206+ cell rate was detected by flow cytometry in PMA‐induced THP‐1 cells co‐cultured with the CM of transfected NSCLC cells. *p < 0.05.
Fig 2: Effects of si‐PTK7 and PIK3CB overexpression on NSCLC cell progression and macrophage polarization. (a) WB was used to evaluate the transfection efficiency of PIK3CB overexpression vector. (b–h) A549 and H1299 cells were transfected with si‐NC/si‐PTK7/vector/PIK3CB. CCK8 assay (b), EdU assay (c), flow cytometry (d,e), and transwell assay (f,g) were performed to assess cell proliferation, apoptosis, and invasion. (h) CD206+ cell rate was evaluated using flow cytometry in PMA‐induced THP‐1 cells co‐cultured with the CM of transfected NSCLC cells. *p < 0.05.
Fig 3: PTK7 interacted with PIK3CB. (a) Heat map showed the differentially expressed genes in NSCLC cells transfected with or without sh‐PTK7 by GSE50138 database. (b) PIK3CB protein expression was analyzed by WB in A549 and H1299 cells transfected with si‐NC/si‐PTK7. (c) TCGA database predicted PIK3CB expression in LUAD tissues and normal tissues. (d) Kaplan‐Meier Plotter database analyzed the relationship between PIK3CB expression and the prognosis of LUAD patients. (e) PIK3CB protein expression was determined by WB in HBE, A549, and H1299 cells. (f) Co‐IP assay was used to confirm the interaction between PTK7 and PIK3CB. *p < 0.05.
Fig 4: Effect of si‐USP8 and PTK7 on NSCLC cell progression and macrophage polarization. (a) The transfection efficiency of PTK7 overexpression vector was confirmed by WB. (b–h) A549 and H1299 cells were transfected with si‐NC/si‐USP8/vector/PTK7. Cell proliferation, apoptosis, and invasion were examined by CCK8 assay (b), EdU assay (c), flow cytometry (d,e), and transwell assay (f,g). (h) Flow cytometry was used to assess CD206+ cell rate in PMA‐induced THP‐1 cells co‐cultured with the CM of transfected NSCLC cells. *p < 0.05.
Fig 5: Effects of sh‐PTK7 and PIK3CB on NSCLC tumor growth. Tumor volume (a) and weight (b) were examined in each group (n = 5). (c) PIK3CB protein level was detected by WB in the tumor tissues of each group (n = 5). (d) IHC staining was performed to measure Ki67, MMP9, and PIK3CB positive cells in the tumor tissues of each group. *p < 0.05.
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