Percentage of positive PBMC after stimulation with PHA in the absence or presence of 3 types of treated MSCs.

<p>After reduction of LLL treated MSCs, CD25+ cells decreased (**P < 0.01 vs. PHA) and CD69+ cells were also reduced compared to control groups (**P < 0.01 vs. PHA).</p

Ser-iPS cells form more teratomas than MEF-iPS cells.

<p>(A) Schematic representation of Ser-iPS cell generation from B6 Sertoli cells by Oct4, Sox2 and Klf4 transfection with and without c-Myc (OSKM or OSK) and teratoma assay in B6 mice. (B) AP staining and analysis of pluripotency markers (Oct4 and SSEA1, red). DAPI staining, blue. Scale bars, 200 µm and 35 µm, respectively. AP staining, Ser-iPS cells (OSKM, clone 1), MEF-iPS cells (OSKM, clone 1) and ES cells (JM8). Oct4 and SSEA1 staining, Ser-iPS cells (OSK, clone 3), MEF-iPS cells (OSK, clone 2) and ES cells (JM8). iPS cell data are representative of all Ser-iPS cells and MEF-iPS cells analyzed. (C) Gene expression in undifferentiated and differentiated Ser-iPS cells (EB assays) determined by qRT-PCR is shown in heatmap format. Expression in undifferentiated ES cells (JM8) was arbitrarily set to 1. Ser-iPS 1 refers to 4F iPS cells (OSKM, clone 1) and Ser-iPS 2 and 3 to 3F iPS cells (OSK, clones 2 and 3); MEF-iPS 1 and 2 refer to 4F and 3F iPS cells (OSKM, clone 1 and OSK, clone 2, respectively). (D) Number and size of teratomas of Ser-iPS cells in B6 mice. B6 MEF-iPS cells and B6 ES cells are shown as controls. Average values of Ser-iPS cells (clones 1, 2 and 3) and MEF-iPS cells (clones 1 and 2) are shown. All Ser-iPS cells and MEF-iPS cells are passage 9–15 (early-passage). *P<0.05, **P<0.01. Bars represent mean ± standard deviation.</p

LLL decrease nuclear translocation of NF-κB.

<p>The images show the cytoplasmic localization of NF-κB in the control cells (upper panel), the nuclear translocation of NF-κB in cells treated with LPS (middle panel) and LLL treatment blocked the nuclear translocation of NF-κB caused by LPS stimulation (lower panel).</p

Primers used in study for PCR.

<p>Primers used in study for PCR.</p

Effect of LLL on mRNA expression and production and of IL-1β, IL4, IL-6, IL-8 and IL-10 by LPS (or not) induced MSCs.

<p>MSCs were incubated with or without LPS (10ng/mL) and simultaneous treated LLL for 1h. Values of *P<0.05, **P<0.01and vs. LPS or control were considered statistically significant.</p

EBs from Ser-iPS cells exhibit reduced CD4 T cell stimulation potential <i>in</i><i>vitro</i>.

<p>(A) Schematic representation of T cell proliferation assay <i>in</i><i>vitro</i>. iPS cells or ES cells were induced to differentiate in EB assays (14 days). EBs were co-cultured with splenic CD4 T cells to assess T cell proliferation. (B) Proliferation of CFSE labeled CD4 T cells co-cultured with Ser-iPS cells (day 12–17) in T cell medium. MEF-iPS cells and ES cells were used as controls. Ser-iPS cells (OSKM, clone 1), MEF-iPS cells (OSKM, clone 1) and ES cells (JM8) of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106110#pone-0106110-g001" target="_blank">Figure 1C</a>. PMA and ionomycin activated T cells, positive control. T cell proliferation refers to the percentage of dividing T cells after 5 days of co-culture. (C) T cell proliferation data after 5 days of co-culture of (B) (n = 3). Average values of Ser-iPS cells (clones 1, 2 and 3) and MEF-iPS cells (clones 1 and 2) are as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106110#pone-0106110-g001" target="_blank">Figure 1D</a>. All Ser-iPS cells and MEF-iPS cells are passage 9–15 (early-passage). *P<0.05; ***P<0.001. Bars represent mean ± standard deviation.</p

LLL suppress ROS and NO promotion induced by LPS in MSCs.

<p>MSCs were incubated with LPS (10ng/mL) and simultaneous treated with LLL for 1 h. Error bars represent the mean±SD (n = 3 per group). Values of **P<0.01 vs. LPS and **P<0.01 vs. control were considered statistically significant.</p

LLL suppress the LPS-induced activation of the NF-κB pathway in MSCs.

<p>Protein samples were analyzed by Western blot using anti p65, p-p65, IκBα and p-IκBα antibody (left), GAPDH was used as the internal control for normalization. The bar chart shows the quantitative evaluation of protein bands by densitometry (right). The data represent the mean±SD (n = 3 per group) *P<0.05, **P<0.01 vs. LPS or control were considered statistically significant.</p