Journal: Scientific reports

Article Title: Protein profiling identified key chemokines that regulate the maintenance of human pluripotent stem cells.

doi: 10.1038/s41598-017-15081-6

Figure Lengend Snippet: Figure 2. Chemokine screening strategy for hPSCs. (a–d) Immunofluorescence of pluripotency markers TRA- 1-81 and Oct4 on hPSCs. KSR medium (a,b); mTeSR1 medium (c,d); hESCs (a,c); hiPSCs (d); Feeder cells (b); Scale bars, 50 μm (a–d; arrows) and 10 μm (insets). Cells were counterstained with DAPI. (e) Chemokine screening strategy for hPSCs cultured in KSR (red) or mTeSR1 (green) media.

Article Snippet: Recombinant human chemokine proteins IL-8, SDF-1α, and IP-10 were purchased from R&D Systems (USA).

Techniques: Immunofluorescence, Cell Culture

Journal: Scientific reports

Article Title: Protein profiling identified key chemokines that regulate the maintenance of human pluripotent stem cells.

doi: 10.1038/s41598-017-15081-6

Figure Lengend Snippet: Figure 4. Chemokine signaling functionally mediates the transmigration of hPSCs. Chemoattractant effect of exogenous IP-10, IL-8, and SDF-1α on hPSCs. hESCs (a,b,e,f,i,j); hiPSCs (c,d,g,h,k,l); IL-8 (a,c) and CXCR2 antagonist SB265610 (b,d); SDF-1α (e,g) and CXCR4 antagonist AMD3100 (f,h); IP-10 (i,k) and CXCR3 antagonist NBI74330 (j,l). Values on graphs represent means ± sem, n = 3 individual experiments. *P < 0.05, **P < 0.01.

Article Snippet: Recombinant human chemokine proteins IL-8, SDF-1α, and IP-10 were purchased from R&D Systems (USA).

Techniques: Transmigration Assay

Journal: Scientific reports

Article Title: Protein profiling identified key chemokines that regulate the maintenance of human pluripotent stem cells.

doi: 10.1038/s41598-017-15081-6

Figure Lengend Snippet: Figure 5. Chemokine IL-8 facilitates the differentiation of hPSCs. Stemness of hPSCs was evaluated by the expressions of three pluripotency genes (Oct4, Nanog, and Rex-1) after treatment with IL-8 and CXCR2 antagonists reparixin/SB265610. hESCs (a–c); hiPSCs (d–f); IL-8 (a,d); Reparixin (b,e); and SB265610 (c,f). Values on graphs represent means ± sem, n = 3 individual experiments. *P < 0.05, **P < 0.01, ***P < 0. 001.

Article Snippet: Recombinant human chemokine proteins IL-8, SDF-1α, and IP-10 were purchased from R&D Systems (USA).

Techniques:

Journal: Scientific reports

Article Title: Protein profiling identified key chemokines that regulate the maintenance of human pluripotent stem cells.

doi: 10.1038/s41598-017-15081-6

Figure Lengend Snippet: Figure 7. Mobility and maintenance of hPSCs require multiple chemokine signals. Schematic representation of predominant chemokines from feeder cells or hPSCs themselves. 1) Released chemokines uniformly mediate the migration of hPSCs; 2) Both differentiating and maintaining chemokines were secreted in the culture supernatant. However, the final effect of chemokines tended to maintain the stemness of hPSCs in vitro.

Article Snippet: Recombinant human chemokine proteins IL-8, SDF-1α, and IP-10 were purchased from R&D Systems (USA).

Techniques: Migration, In Vitro

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammation-induced chondrocyte hypertrophy is driven by receptor for advanced glycation end products.

doi: 10.4049/jimmunol.175.12.8296

Figure Lengend Snippet: FIGURE 5. sRAGE and anti-RAGE inhibit CXCL8- and TNF-- but not ATRA-induced type X collagen expression. Primary normal human knee articular chondrocytes (donor ages, 29–62) (5 105 cells/six-well dish) were stimulated with CXCL8 (A), ATRA (B), or TNF- (C) in the presence or absence of 1 g/ml sRAGE or 20 g/ml anti-RAGE. SDS- PAGE and Western blotting analysis for type X collagen were performed on cell lysates at 5 days in culture. These data are representative of results from six normal human donors.

Article Snippet: All-trans retinoic acid (ATRA), and human recombinant CXCL8 and TNF- were from R&D Systems, and rabbit polyclonal Abs to type X collagen were from Calbiochem. mAb to RAGE was from Chemicon International.

Techniques: Expressing, SDS Page, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammation-induced chondrocyte hypertrophy is driven by receptor for advanced glycation end products.

doi: 10.4049/jimmunol.175.12.8296

Figure Lengend Snippet: FIGURE 3. RAGE expression and inducible S100A11 secretion in cul- tured human articular chondrocytes. A, RAGE expression. First-passage human knee articular chondrocytes (5 105 cells/six-well dish) from six normal donors (age range, 29–62), were prepared as described in Mate- rials and Methods, were studied for RAGE expression by flow cytometric analysis. The panels, taken from studies of one normal donor representative of all the donors, depict binding of isotype control IgG (open histogram) and additionally of anti-RAGE IgG without stimulation or in response to CXCL8, TNF-, and ATRA for 24 h (solid histograms). In normal chon- drocytes, constitutive RAGE expression was detected in 59 9% of cells, and there were no significant changes in the proportions of RAGE-express- ing chondrocytes in response to CXCL8, TNF-, or ATRA, as seen in the representative results from the single donor demonstrated here. B, S100A11 expression and multimerization in chondrocytes. SDS-PAGE/ Western blotting analysis was performed on 1 g of human recombinant S100A11 isolated from transfected 293 cells as described in Materials and Methods (left side of figure), and on 30-g aliquots of protein precipitated from conditioned medium of first-passage normal human knee chondro- cytes (right side of figure) (donor ages, 29–62). The chondrocytes were prepared under the same conditions as for A above, and stimulated for 24 h by addition of S100A11, CXCL8, TNF-, or ATRA, at which time con- ditioned media were studied. Results are representative of those from six different normal human donors, as in A.

Article Snippet: All-trans retinoic acid (ATRA), and human recombinant CXCL8 and TNF- were from R&D Systems, and rabbit polyclonal Abs to type X collagen were from Calbiochem. mAb to RAGE was from Chemicon International.

Techniques: Expressing, Binding Assay, Control, SDS Page, Western Blot, Recombinant, Isolation, Transfection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammation-induced chondrocyte hypertrophy is driven by receptor for advanced glycation end products.

doi: 10.4049/jimmunol.175.12.8296

Figure Lengend Snippet: FIGURE 7. Schematic of RAGE signaling in the induction of chondro- cyte hypertrophy. This schematic depicts the induction by CXCL8 and TNF- of secretion of S100A11 (and likely other chondrocyte-expressed RAGE ligands not limited to calgranulins). The paradigm further depicts subsequent RAGE signaling critically transduced by p38 MAPK pathway activation as a central event in chondrocyte hypertrophy associated with low-grade inflammation in the OA joint articular cartilage.

Article Snippet: All-trans retinoic acid (ATRA), and human recombinant CXCL8 and TNF- were from R&D Systems, and rabbit polyclonal Abs to type X collagen were from Calbiochem. mAb to RAGE was from Chemicon International.

Techniques: Activation Assay