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. 2013 Nov 20;8(11):e79458. doi: 10.1371/journal.pone.0079458

Figure 4. SEL1L affects cell adhesion.

Figure 4

(A) MIN6 cells were transfected with: scrambled-control (white bars), siRNA against Sel1l (black bars) or co-transfected with Sel1l-siRNA and β1 integrin over-expressing construct (grey bars) were left to adhere for 1 hour on 96-well plate coated with Collagen-IV (CollIV), Fibronectin (FN), Laminin (LN) and Vitronectin (VN). Adherent cells were then fixed, stained and counted under a microscope. Values are relative to untransfected adherent cell and presented as means ± SD from three independent experiments. (B) Western blot analysis of MIN6 cells (NT) transfected with scramble-control (Scr), with siRNA against Sel1l (iSel1l) and co-transfected with β1 integrin over-expressing construct (Scr+Itgb1and iSel1l+Itgb1) probed for SEL1L, β1 integrin (ITGB1), phospho-ERK1/2 and total-ERK1//2. (C) Quantification of the immunoblot bands; values are representative of three independent experiments and are expressed as fold of expression ± SD relative to NT and normalized to total ERK1/2 as loading control. (D) Photomicrographs showing the effect of SEL1L down-modulation on MIN6 morphology; the Sel1l-interfered cells (iSel1l) appears round-shaped compared to controls (Scr, NT, Scr+Itgb1) while overexpression of β1 integrin subunit restore MIN6 polygonal spindle-like morphology (iSel1l+Itgb1).