Figure 5

Integrins are important regulators of the self-renewal capacity of ES-D3 cells. A) Control (Ctrl), Itgα6-, ItgαV- and Itgβ1-KD ES-D3 cells (300 cells/mm²) were seeded onto LN-511-, Col-I- or FN-coated tissue culture dishes (3.5 cm Ø) and cultured in the absence or presence of 10 ng/ml of LIF. Upon confluency the cells were trypsinized, counted and reseeded at 300 cells/mm². Cell doubling index was calculated for each time point as described in materials and methods. The graph represents data from 2 independent experiments performed in duplicates. Cross (†) indicates samples where sufficient amount of cells could not be harvested for analysis. B) Control, Itgα6-, ItgαV- and Itgβ1-KD ES-D3 cells were grown on LN-511-, Col-I- and FN-coated surfaces in the absence of LIF, total RNA was extracted at day 5, passage 3 (P3) and P5 and the mRNA expression levels of self-renewal markers (Nanog, Sox2, and Oct3/4), ES cell markers (Rex1, Klf4 and Tbx3) and a differentiation marker (FGF5) were analyzed by qPCR. The data is shown as heat maps where red indicates downregulation of the mRNA and green indicates up-regulation of the respective mRNAs compared to ES-D3 cells grown on LN-511 in the presence of LIF on day 5. The raw data values of means +/−STD are shown in Figure S1 (see Additional file 3). The graph represents data from 2 independent experiments performed in duplicates. Gray color indicates no data due to loss of cells during culture.