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. 2022 May 24;23(11):5874. doi: 10.3390/ijms23115874

Figure 5.

Figure 5

Effect of integrin αvβ3 blockade and ZEB1 knockdown on the SPARC-induced E-cadherin downregulation and enhanced migration (A). Representative Western blot of ZEB1, E-cadherin and SPARC in LNCaP cells transduced with a lentiviral vector carrying a short hairpin RNA against ZEB1 (shZEB1) or a scrambled sequence (shScr) (B). Quantification of the optic density of the Western blot shown in (A). Protein expression normalized to β-actin and control cells (shSCR) (C,D). Relative expression of E-cadherin measured by RT-qPCR in LNCaP shScramble (C) and LNCaP shZEB1 cells stimulated with 1 μg/mL SPARC and 50 μM RGD for 6 h. E-cadherin expression normalized to pumilio and control cells (first column) using the ΔΔCt method (E,F). Transwell migration assay of LNCaP shScramble (E) and LNCaP shZEB1 (F) cells stimulated with 1 μg/mL SPARC and 50 μM RGD for 24 h (G,H). Representative images of wound healing assay of LNCaP shScramble (G) and LNCaP shZEB1 (H) cells stimulated with 1 μg/mL SPARC and 50 μM RGD for 24 h (I,J). Quantification of the percentage of the wound area covered after 24 h (B–F,I,J). Relative migrations were compared with their own control (basal condition) (B–F). Data are expressed as mean ± SD (n = 3). ns = p > 0.05; * = p ≤ 0.05; ** = p ≤ 0.01; *** = p ≤ 0.001; (B)—Mann–Whitney U test; (C–F,I,J)—Kruskal–Wallis test.