Figure 6.
OSBP is highly expressed in epithelial cells. (A and B) EMT was induced in MDCK cells stably expressing P4M-SidM-EGFP by HGF treatment (A). TGF-β was used to induce EMT in A549 cells (B), and then cells were transfected with P4M-SidM-EGFP plasmid. Cells were mounted for epifluorescent live cell imaging to asses OSBP-dependent PI(4)P consumption. OSW-1 was added to the cells at the indicated time points after the start of imaging. Intensities in Golgi areas were measured to plot the kinetics of the P4M-SidM-EGFP experiments. N = number of independent experiments presented, n = number of cells analyzed. Each line corresponds to one cell and bold lines with the shaded area indicate mean ± 95% confidence interval. P values were calculated by two-way ANOVA followed by Sidak’s multiple comparison test. The three independent experiments are shown in Fig. S5, A and B. Bar = 10 μm. (C) Expression heatmap of epithelial markers, mesenchymal markers and ORP genes across cell lines of the Cancer Cell Line Encyclopedia collection. (D) Using the marker gene expression data, epithelial-mesenchymal indexes were calculated for each cell line and ranked. (E) Epithelial-mesenchymal indexes were correlated with ORP expressions in each cell line then Pearson r values were ranked to reveal ORPs associated with epithelial or mesenchymal cells. OSBPL2 and OSBP are epithelial, while OSBPL6 and OSBPL8 are mesenchymal-specific ORPs. (F) EMT was triggered in A549 cells with 72 h TGF-β treatment, then RT-qPCR was performed to assess expression changes in EMT-transcription factors and selected ORPs. Means are calculated from three independent experiments. *P < 0.05; **P < 0.01; ****P < 0.001; ns = non-significant, Welch’s t test. (G and H) OSBP and OSBPL2 expression data was obtained from TCGA Lung adenocarcinoma samples (n = 719) and patients were split according to median gene expression values to obtain high and low expression groups. Low OSBP and OSBPL2 expressions are associated with a poor clinical outcome in lung adenocarcinoma patients. P values were calculated by Gehan–Breslow–Wilcoxon test.

OSBP is highly expressed in epithelial cells. (A and B) EMT was induced in MDCK cells stably expressing P4M-SidM-EGFP by HGF treatment (A). TGF-β was used to induce EMT in A549 cells (B), and then cells were transfected with P4M-SidM-EGFP plasmid. Cells were mounted for epifluorescent live cell imaging to asses OSBP-dependent PI(4)P consumption. OSW-1 was added to the cells at the indicated time points after the start of imaging. Intensities in Golgi areas were measured to plot the kinetics of the P4M-SidM-EGFP experiments. N = number of independent experiments presented, n = number of cells analyzed. Each line corresponds to one cell and bold lines with the shaded area indicate mean ± 95% confidence interval. P values were calculated by two-way ANOVA followed by Sidak’s multiple comparison test. The three independent experiments are shown in Fig. S5, A and B. Bar = 10 μm. (C) Expression heatmap of epithelial markers, mesenchymal markers and ORP genes across cell lines of the Cancer Cell Line Encyclopedia collection. (D) Using the marker gene expression data, epithelial-mesenchymal indexes were calculated for each cell line and ranked. (E) Epithelial-mesenchymal indexes were correlated with ORP expressions in each cell line then Pearson r values were ranked to reveal ORPs associated with epithelial or mesenchymal cells. OSBPL2 and OSBP are epithelial, while OSBPL6 and OSBPL8 are mesenchymal-specific ORPs. (F) EMT was triggered in A549 cells with 72 h TGF-β treatment, then RT-qPCR was performed to assess expression changes in EMT-transcription factors and selected ORPs. Means are calculated from three independent experiments. *P < 0.05; **P < 0.01; ****P < 0.001; ns = non-significant, Welch’s t test. (G and H) OSBP and OSBPL2 expression data was obtained from TCGA Lung adenocarcinoma samples (n = 719) and patients were split according to median gene expression values to obtain high and low expression groups. Low OSBP and OSBPL2 expressions are associated with a poor clinical outcome in lung adenocarcinoma patients. P values were calculated by Gehan–Breslow–Wilcoxon test.

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