Tlx1-related gene expression analysis.

<p><b>(A)</b> Tlx1 (Hox11)-related gene expression levels were significantly altered. A hierarchical clustering image reveals differences between Samples 1, 2, and 3. <b>(B)</b> Photomicrographs of transplanted islet cells in the spleen on days 0, 154, and 290. Sections were stained with anti-insulin antibody (green) and anti-Rrm2b antibody (red). Scale bars: 100 μm. The yellow boxed regions in the second column were enlarged, and the scale bars are 200 μm. <b>(C)</b> Photomicrographs of transplanted islet cells in the spleen on days 0, 154, and 290. Sections were stained with anti-insulin antibody (green) and anti-Pla2g2d antibody (red). Scale bars: 100 μm. The yellow boxed regions in the second column were enlarged, and the scale bars are 200 μm.</p

Recovery of blood flow in ischemic limbs treated with M-Mø and GM-Mø.

<p>Representative laser Doppler perfusion images were taken at indicated intervals for the PBS control group, M-Mø group, and GM-Mø group. Data are expressed as mean ± SEM, n = 5 group, **<i>P</i><0.01, *<i>P</i><0.05.</p

Flow cytometric analysis of F4/80⁺ macrophages from M-CSF- or GM-CSF- cultured BMCs.

<p>Data is representative of three independent experiments.</p

Early inflammatory reactions after receiving the marginal number of islets into the liver, beneath the kidney capsules, or into the spleen.

<p><b>(A–C)</b> Plasma MCP-1, G-CSF, and HMGB1 levels were measured 6 hours after islet transplantation into the portal vein (PV), beneath the kidney capsule (KC), or into the spleen (SP) (n = 7). Untreated naïve mice were used as a control (n = 7). Values are means±SD. *p<0.05. <b>(D)</b> Photomicrographs of islet cells after transplantation in the PV, KC, or SP. Sections were stained with anti-Gr-1 or F4/80 followed by staining with Haematoxylin. Scale bars: 100 μm.</p

Expression of cytokine mRNA, and IL-10 protein levels in M-CSF- or GM-CSF-cultured BMCs.

<p>(A) The expression of proinflammatory cytokines (TNF-alpha, IL-6, IL-1beta, IL-12a, and IL12b) and anti-inflammatory cytokine (IL-10) mRNA in M-CSF- or GM-CSF-cultured BMCs. (B) IL-10 concentration in the supernatant of M-CSF- or GM-CSF-cultured BMCs. Data are expressed as mean ± SEM, ***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05.</p

Long-term effects of intra-splenic islet transplantation.

<p><b>(A)</b> Non-fasting blood glucose levels in STZ-induced diabetic mice (C57BL/6) transplanted with 25 syngeneic islets in the spleen (SP) and 100 syngeneic islets under the kidney capsule (KC). Individual lines represent glucose levels in each animal. <b>(B)</b> Photomicrographs of transplanted islet cells in the SP on day 290. Sections were stained with anti-insulin antibody followed by hematoxylin. Scale bars: 100 μm. <b>(C)</b> Insulin content was measured after islet transplantation on days 0 (n = 6) and 280 (n = 6). Values are means±SD. *p<0.0001.</p

Recovery of blood flow in ischemic limbs treated with M-CSF-cultured BMCs and GM-CSF-cultured BMCs.

<p>Representative laser Doppler perfusion images taken at indicated intervals for the PBS control group, M-CSF-cultured BMCs group and GM-CSF-cultured BMC group. Data are expressed as mean ± SEM, n = 5–7/group, **<i>P</i><0.01, *<i>P</i><0.05.</p

Foxp3 expression in M-Mø or GM-Mø treated ischemic hind limbs and the recovery of blood flow in ischemic limbs treated with IL-10.

<p>(A) Photomicrographs show representative DAB staining for Foxp3 (scale bar = 50 µm). The number of Foxp3 positive cells was counted in five random images for each mouse. The thigh muscle of each group was isolated 7 days after the induction of ischemia. (B) IL-10 levels in thigh muscle after limb ischemia at 7 days after intramuscular injection of indicated cells (data are expressed as mean ± SEM, n = 3 group, ***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05). (C) Recovery of blood flow in ischemic limbs treated IL-10. Representative laser doppler perfusion images were taken at indicated intervals for the PBS control groups and IL-10 treated groups. Data are expressed as mean ± SEM, n = 5–7/group.</p

vWF and LYVE-1 staining in M-Mø and GM-Mø treated ischemic hind limbs.

<p>Representative photomicrographs (original magnification ×400). vWF positive and LYVE-1 positive cells are stained green and red, respectively (***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05).</p

The presence of DNA methylation profiles specific to individual types of mouse leukocytes.

(A) Unsupervised hierarchical clustering analysis using the top 1% (2,588 probes) of the total 258,525 probes that had the HSD. The three myeloid lineage cell types and four lymphoid lineage cell types were clustered separately. (B) Origins of the 2,588 probes against gene structure. 291 (11.3%) and 283 (10.9%) of probes were derived from enhancers and TSS200, respectively. (C) Origins of the 291 enhancer and 283 TSS200 probes against CGI. The vast majority of the HSD probes were in open sea, and 0 enhancer probe and 1 TSS200 probe were in CGI. (D) Unsupervised hierarchical clustering analysis using the top 1% (155 probes) from enhancers. The three myeloid lineages and the four lymphoid lineages were classified, similar to the use of the HSD probes from all genomic regions. (E) Unsupervised hierarchical clustering analysis using the top 1% (95 probes) from TSS200CGIs. Separation of the two lineages was not clear. (F) and (G) Principal component analysis (PCA) using the HSD probes from enhancers (F) and those from TSS200 (G). Separation of the two lineages was clearer using probes from enhancers.</p