BMP-2 and MMP-2 expression in ARPE-19 cells at different times points following ATRA treatment.

ATRA at 10−6 mol/l upregulated the expression of BMP-2 mRNA in ARPE-19 cells, and this effect was time dependent. There were no significant changes in BMP-2 mRNA in ARPE-19 cells after incubation with ATRA for 12 h, but the mRNA levels of BMP-2 were significantly increased after treatment with 10−6 mol/l ATRA for 48 h and 72 h (p−6 mol/l for 6 h, peaked at 12 h and 24 h (P0.05) (Fig 4B). The expression levels of BMP-2 and MMP-2 protein were similar to the levels of those mRNA in ARPE-19 cells (Fig 4A and 4B). Cell supernatant from the cells treated with ATRA concentrations of 10−6 mol/l were analyzed by ELISA and compared with controls (medium with 0.1% DMSO). ATRA also upregulated the BMP-2 protein level in a time dependent manner. BMP-2 protein expression was significantly increased after the cells were treated with 10−6 mol/l ATRA at both 48 h and 72 h (p<0.05) (Fig 4C). The MMP-2 protein level was similar to its mRNA level and was significantly increased at 12 h (P<0.05) (Fig 4D).</p

LE135 could block the pathway of RARβ in ARPE-19 cells.

Compared to the control, LE135 (10-6mol/l) could block the ATRA-induced increase in RARβ expression at either the mRNA (P<0.05) (Fig 5A) or the protein level (P<0.001) (Fig 5B and 5C) at 48 h.</p

The Effect of ATRA on survival in ARPE-19 cells.

ATRA caused a low level of apoptosis (−9 to 10−6 mol/l, but the cell morphology remained normal. The cells were inhibited at the highest concentration of ATRA treatment 10−5 mol/l (Fig 1A). At both 24 h and 48 h, the cell proliferation was inhibited by ATRA in a dose-dependent manner. A small reduction (−6 mol/l, but these changes were not statistically significant (P>0.05). When the ATRA concentration was 10−5 mol/l, the cell survival rate was below 50% at both time points (Fig 1B).</p

PBDTTTZ: A Broad Band Gap Conjugated Polymer with High Photovoltaic Performance in Polymer Solar Cells

PBDTTTZ: A Broad Band Gap Conjugated Polymer with High Photovoltaic Performance in Polymer Solar Cell

Primer sequences used in RT-PCR.

<p>Primer sequences used in RT-PCR.</p

ATRA-induced BMP-2 and MMP-2 expression were inhibited by RARβ.

<p>The increased expression of BMP-2 and MMP-2 induced by ATRA were almost completely blocked by pre-treatment of the ARPE19 cells with LE135. ATRA at 10⁻⁶ mol/l upregulated the expression of BMP-2 and MMP-2 on both mRNA and protein levels in ARPE-19 cells (P<0.05), but there were no significant changes after incubation with LE135 (10^-6mol/l) or ATRA (10^-6mol/l) added to LE135 (10^-6mol/l) (Fig 6A, 6B and 6C).</p

Time-related changes in expression of RARβ mRNA and protein.

<p>A significant increase in the RARβ band intensity was detected for both mRNA (Fig 3A and 3B) and protein (Fig 3A and 3C) at 24 h and 48 h after treatment with 10⁻⁶ mol/l ATRA (P<0.05).</p

RARs and RXRs expressions in ARPE-19 cells.

<p>RARs mRNA was strongly expressed in ARPE-19 cells, whereas RXRs expression was not detected (Fig 2A and 2B). RARβ mRNA levels was increased after ATRA treatment when compared with controls. Expression of RARβ mRNA was dose dependent with a maximal effect observed with 10⁻⁵ mol/l ATRA. In contrast, there were no significant changes in the mRNA levels of RARα and RARγ (P>0.05) (Fig 2C and Fig 2D).</p

Additional file 1 of Refractive error characteristics and influence on ocular parameters in patients with unilateral congenital ptosis

Additional file 1

Primer sequences used in qRT-PCR.

<p>Primer sequences used in qRT-PCR.</p