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1.
Figure 2

Figure 2. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

Integrin expression profiles in the Organ of Corti. RT-PCR of RNA isolated from organ of Corti of adult mice. Bands at the predicted molecular weight are visible for integrin β1, β3, αv, α6 and β-actin but not for integrin β4. cDNA was omitted for the negative control.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
2.
Figure 7

Figure 7. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

Graph of changes in expression levels of integrin subunits and myosin VIIa during OC-2 cell differentiation. Schematic time course graph showing fluorescence values of αv-, α6-, β1- and β3- integrin subunit surface expression measured by FACS and chemiluminescence values of the hair cell marker myosin VIIa in OC-2 cells measured by Western blot, over the 14 day differentiation process.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
3.
Figure 3

Figure 3. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

αv-integrin expression profiles change during differentiation. Expression of (A) α6-integrin and (B) αv-integrin were examined by FACS at days 2, 6, 10 and 14 during the differentiation process. The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). α6-integrin expression profiles did not change throughout the time course. αv-integrin expression profiles show an increase at day 6 and are at comparable levels with fully differentiated OC-2 cells at day 14 with an apparent even higher level at day 10.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
4.
Figure 6

Figure 6. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

Effect of β3-integrin-siRNA treatment on integrin surface expression. Differentiated OC2 cells were treated with β3-siRNA and assayed by FACS for expression of surface integrins. The data is compared to integrin expression levels in normal differentiated cells (39°), differentiated cells exposed to scrambled siRNA sequences (siRNAβ3) and undifferentiated cells (33°). Exposure to β3-integrin-siRNA resulted in a significant reduction of the surface expression of β3, β1- and αv- to levels similar to those of undifferentiated cells. There was no significant effect of scrambled siRNA sequences on any integrin expression and β3-integrin-siRNA did not significantly affect α6-integrin providing further evidence for the specificity of the siRNA inhibition.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
5.
Figure 4

Figure 4. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

β1 and β3-integrin expression profiles change during differentiation. Surface expression of (A) β1-integrin and (B) β3-integrin was examined by FACS at days 2, 6, 10 and 14 of the differentiation process The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). Increase of β1-integrin expression profile is visible from day 2 and it reaches the level of differentiated cells by day 10. β3-integrin expression which is undetectable above the Ig signal up to day 2, becomes visible at day 6 and gradually reaches the differentiated levels at day 14.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
6.
Figure 5

Figure 5. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

β3-integrin is a marker for OC-2 cell differentiation. (A) Western blot analysis of myosin VIIa expression in 39°C OC-2 cells untreated or after treatment with either scrambled (Scr), β1-, α6-, αv- and β3-integrin siRNA. Treatment with all siRNAs reduced myosin VIIa levels significantly when compared with untreated OC-2 cells or OC-2 cells treated with scrambled siRNA. (B) Undifferentiated OC-2 cells transduced with human β3-integrin (+hβ3) had similar levels of surface β3-integrin to differentiated (39) OC-2 cells. Furthermore, analysis of the expression of myosin VIIa showed it was increased significantly in OC-2 cells transduced with human β3-integrin (+hβ3) cells compared with undifferentiated (33) cells. Bar graphs represent densitometric results of mean relative values of myosin VIIa levels ± s.e.m. HSC-70 was used as a loading control. *P < 0.05, **P < 0.01. n = 3 individual experiments.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.
7.
Figure 1

Figure 1. From: ?3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear.

Integrin expression profiles are altered after OC-2 cell differentiation. (A) Western blot analysis of myosin VI and myosin VIIa levels in OC-2 cells grown at 33°C and 39°C. Both myosin VI and VIIa levels were increased significantly in cells grown at 39°C cells in comparison with those incubated at 33°C. Bar graphs represent densitometric results of mean relative values of myosin VI and myosin VIIa levels ± s.e.m. HSC-70 provided the loading control. (B) Undifferentiated (33°C) and differentiated (39°C) OC-2 cells were analysed by FACS for the integrin subunits α6, β1, αv and β3. Levels of β1-, αv- and β3-integrin subunits were increased significantly in differentiated OC-2 cells when compared with undifferentiated OC-2 cells. α6-integrin surface expressions levels did not change between the two cell phenotypes. Bar graph represents mean fluorescence units of the various integrin subunits ± s.e.m.; n = 3 independent experiments. White bars = cells at 33°C, black bars = cells at 39°C cells; nd = no significant difference, *P < 0.05, **P < 0.01.

Ivan Brunetta, et al. BMC Cell Biol. 2012;13:5-5.

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