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. 2021 Nov 20;298(1):101440. doi: 10.1016/j.jbc.2021.101440

Figure 1.

Figure 1

Tumorigenic role of ARHGEF17.A and B, NSCLC TCGA patients with ARHGEF17 amplification (GEF17; AMP) compared with WT according to their tumor stage (A, stages I–IV) and lymph node disease (B, N0-3,NX) were analyzed by Chi-squared test at the cBioportal platform. C, maximally separated Kaplan–Meier plots of NSCLC (TCGA datasets) compared the survival of patients with high and low GEF17 expression (stages II–IV). D, GEF17 knockdown in LAP0297 cells. A representative Western blot is shown. Graph represents mean ± SEM, n = 3. ∗∗p < 0.01; t test. E, LAP0297 tumor growth in immunocompetent FVB mice inoculated with ARHGEF17-knockdown (sh-GEF17) or control cells (sh-Control). Graph represents the mean ± SEM; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; ANOVA followed by Tukey's test, n = 10. F, tumor weight was measured at day 15. Graph represents the mean ± SEM; ∗p < 0.05; t test. n = 10. G, representative tumors are shown. The scale represents 1 cm. H, lung weight. Graph shows the mean ± SEM weight of normal lungs from healthy mice and metastasized lungs from mice inoculated in the tail vein with sh-Control or sh1-GEF17 LAP0297 cells. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; Mann–Whitney U test. n = 6 control groups, n = 7 sh-GEF17. The scale represents 1 cm. I, macrometastasis in the lungs of FVB mice inoculated in the tail vein with sh-Control or sh1-GEF17 LAP0297 cells. Graph represents the number of superficial metastases, mean ± SEM; ∗p < 0.05; t test. n = 6 sh-Control, n = 7 sh-GEF17. Normal lungs from healthy mice are shown at the left. The scale represents 3 mm, zoom: 1 mm. J and K, in vitro proliferation in response to 10% FBS (J) and viability in cells incubated in serum-free media for 48 h (K) were assessed with sh-Control and shGEF17 LAP0297 cells with the MTT assay. Fifteen hundred cells per well were left overnight in 1% FBS for proliferation assays and 50,000 cells for viability experiments, followed by 48 h in media containing 10% FBS (J) or in serum-free media (K). Proliferation and viability were evaluated with the MTT assay; in both cases, cells incubated in 1% FBS were used as a control. Graphs represent the mean ± SEM. J, n = 4: ∗∗∗p < 0.001, ∗∗p < 0.01, ns; t test. K, n = 4; ∗p < 0.05, ns; t test. MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; ns, nonsignificant; NSCLC, non–small cell lung carcinoma.