Page 67 - Physico-Chemical Niche Conditions for Bone Cells

Figure 5. Effect of 1 h PFF on integrin-a5 expression and distribution in MC3T3-E1 osteoblasts. (A) mRNA expression of ITGA51. (B) 2D images of control cells stained for integrin-a5 (green) and DAPI for the nucleus (blue). (C) 2D images of 1 h PFF-treated cells stained for integrin-a5 (green) and nucleus (blue). Bar = 50 μm. (D) Fluorescence intensity of integrin-a5 in control and PFF-treated cells. (E) Integrin-a5 number in control and PFF-treated cells. (F) Integrin-a5 area in control and PFF-treated cells. (G) The integrin-a5 average size in control and PFF-treated cells. * p < 0.05. CON, control; PFF, pulsating fluid flow.
Expression of a-Tubulin and Nesprin 4
a-Tubulin immunofluorescence staining of microtubules indicated that microtubules were similarly distributed in control and PFF-treated cells, with microtubules centered around the nuclei, extending in all directions (Figure 6). The distribution of the microtubule staining intensity was similar between PFF-treated and static cultured cells, i.e., microtubule staining intensity was uniform when comparing the perinuclear area vs. the cell periphery (Figure 6). a-Tubulin protein expression levels in MC3T3-E1 osteoblasts, either or not treated with PFF, were assessed by western blot analysis (Figure 6). a-Tubulin protein expression was similar after static and PFF-treatment (Figure 6; p = 0.698, n = 4). Nesprin 4 gene expression was significantly down-regulated by PFF treatment (Figure 6).
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