The morphology of the BMSCs after co-culturing.

<p>A, The morphological changes of BMSCs over 7 days (10×); B, The bar graph demonstrates that the percentage of neural-like BMSCs increases when co-cultured with RGCs. p≤0.05 at all time points. (10 pictures were taken randomly under a regular microscope from culturing cells at different time points. The average relative percent of neural-like BMSCs come from at least three separate experiments).</p

Patterns of differentially expressed genes in BMSCs.

<p>A, 5% polyacrylamide gel eletrophoresis demonstrates two dense bands representing upregulated genes in BMSCs co-cultured with RGC (white arrowheads). B, RT-PCR using total RNA isolated from BMSCs on day three demonstrated increased TSP-1 mRNA expression compared to controls. C, BMSC TSP-1 protein expression was increased as compared to the controls as demonstrated by Western blot. β-actin was used as a loading control in B and C. D, TSP-1, TSP-2 and TSP-1 receptors (CD36, CD47) expression were determined by real-time RT-PCR. E, Western analysis for TSP-1 in co-culture medium. *p-values≤0.05. (All data come from at least three separate experiments).</p

Endogenous TSP-1 expression was silenced by siRNA and rescued by infection with a recombinant adenovirus containing the wild-type TSP-1 cDNA driven by the CMV promoter (pAd/CMV-Myc-TSP-1).

<p>(The experiments were repeated at least three times).</p

Morphological changes of BMSCs after co-culturing with RGCs when TSP-1 was silenced with siRNA and rescued by infection with a recombinant adenovirus expressing TPS1.

<p>48 hours after the first siRNA transfection, BMSCs and RGCs were co-cultured in a transwell system. A, Neural-like morphological changes (10×) in BMSCs was increased at 3 and 5 days in controls (1). This effect was diminished when TSP-1 was silenced with siRNA (2), but restored in BMSCs infected with a TSP-1-expressing adenovirus (3). B, Bar graph shows the mean and SE of the percentage of neural-like cells in the control, silenced, and rescued BMSCs (p = 0.011, p = 0.022, p = 0.028, respectively). *p-values≤0.05. (10 pictures were taken randomly under a regular microscope from culturing cells at different time points. The average relative percent of neural-like BMSCs come from at least three separate experiments).</p

The primers Real-time RT-PCR.

<p>The primers Real-time RT-PCR.</p

The effect of BMSCs on RGCs.

<p>A, RGC neurite formation was enhanced after 5 days of co-culturing with BMSCs (10×). B, Synaptophysin, MAP-2 and TGF-β expression were up-regulated in RGCs co-cultured with BMSCs as determined by real-time RT-PCR (p = 0.031, p = 0.023, p = 0.015, respectively). C, RGC survival was increased when co-cultured with BMSCs (p = 0.025) (d) The mean outgrowth of neurites in RGCs co-cultured with BMSCs increased (p = 0.03). *p-values≤0.05. (10 pictures were taken randomly under a regular microscope from culturing cells at different time points. The average relative percent of surviving and outgrowth of RGCs come from at least three separate experiments).</p