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Figure 2 | BMC Cell Biology

Figure 2

From: Regulation of the microtubular cytoskeleton by Polycystin-1 favors focal adhesions turnover to modulate cell adhesion and migration

Figure 2

Polycystin-1-dependent migration depends on both actin and microtubules, while front-rear polarity only on microtubules. (A) Boyden chambers on mIMCD cells, showed that silencing of Pkd1 (mIMCDshPkd1: clones C12, C16) decreases their migration capability, as compared to controls (mIMCDshCtrl: clones M3, M4). (B) Golgi orientation in wound-healing assays on mIMCD cells, showed front-rear polarity impairment upon Pkd1 silencing. (C) Boyden chambers assay performed on MDCK cells (left panel) and fibroblasts (right panel) in the presence of CytochalasinD (cyt) or LatrunculinA (lat), revealed the essential role of actin in migration. Graphs are representative of three independent experiments; averages and SD are shown. (D) MDCKZeo (clone F6) and MDCKPKD1Zeo (clone 68) were wounded in the presence of CytochalasinD (cyt) or LatrunculinA (lat) and after 3 h stained with phalloidin (actin), anti-giantin (Golgi) and DAPI (nucleus). Bar: 50 μm. (E-F) Quantification of Golgi repositioning revealed no effect of CytochalasinD (E, cyt) or LatrunculinA (F, lat) on front-rear polarity in MDCKPKD1Zeo cells (clones 36, 68, G3). Graphs are representative of three independent experiments; averages and SD are shown. (G) Boyden chamber assays performed in the presence of Nocodazole (noc) or Taxol (tax) revealed the role of microtubules in migration. Graph is representative of three independent experiments; averages and SD are shown. (H-I) Quantification of Golgi repositioning revealed a strong effect of Nocodazole and Taxol on front-rear polarity in MDCKPKD1Zeo epithelial cells (H) and in Pkd1 +/+ (I). Graphs are representative of three independent experiments; averages and SD are shown. (L) Front-rear polarity was evaluated as in D in MDCKZeo (clone F6) and MDCKPKD1Zeo (clone 68) cells treated with Nocodazole and Taxol. Bar: 50 μm. For all graphs in this figure: Statistical analysis ANOVA; NS non-statistically significant (p > 0.05), *p < 0.05, **p < 0.01, ***p < 0.001, referred to the relative control (ct).

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