Fig. 4: Proliferation, migration and invasion in WT, TPC2 KO and Rab7a KO SK-MEL-5 cells. | Nature Communications

Fig. 4: Proliferation, migration and invasion in WT, TPC2 KO and Rab7a KO SK-MEL-5 cells.

From: Rab7a is an enhancer of TPC2 activity regulating melanoma progression through modulation of the GSK3β/β-Catenin/MITF-axis

Fig. 4

Ctb assay assessing proliferation of SK-MEL-5 cells monitored over 96 h comparing WT cells to different clones for TPC2 KO (n = 12) (a) and Rab7a KO (n = 18) (f). Genetic ablation of either TPC2 (b) or Rab7a (g) in SK-MEL-5 melanoma line shows significantly slower invasion and migration, cells seeded on transwell chambers and monitored overnight. Statistical analysis for Boyden chamber migration and invasion experiments in TPC2 KO (n = 11–34) (c) and Rab7a KO (n = 13–49) (h) SK-MEL-5 cells. Clonogenic assay showing significant reduction in survival and growth as single colonies for both TPC2 KO (d) and Rab7a KO (i) SK-MEL-5 cells. Statistical analysis for SK-MEL-5 TPC2 KO (n = 6) (e) and Rab7a KO (n = 6) (j) plotted as colony area percentage, fold induction on WT cells. k qPCR (n = 2) and Western blot (n = 8) analysis indicating unchanged transcript and protein levels of Rab7a in TPC2 KO clones. Statistical significance in a and f was carried out using two-way ANOVA followed by Bonferroni multiple comparisons test (n = 3, each), in c by one-way ANOVA, and in e, h, and j by two-tailed unpaired t-test Student’s t-test. Shown are mean values ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All scale bars = 25 µm. All n numbers represent biological replicates. Source data are provided as Source Data file.

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