Journal: Nano-Micro Letters

Article Title: Dual-Wavelength Photosensitive Nano-in-Micro Scaffold Regulates Innate and Adaptive Immune Responses for Osteogenesis

doi: 10.1007/s40820-020-00540-z

Figure Lengend Snippet: a Schematic diagram of biomaterial-mediated DC maturation and T cell activation. b FACS of all DCs (CD11c + , IA/IE +) and mature DCs (CD11c + , IA/IE + , CD86 +) recruitment at 2 weeks after biomaterials implantation in vivo. c Semiquantification of gating strategy cells in (B). n = 6, **** P < 0.0001. d IHC staining of CD11c—DC marker and CD83/CD40—mature DC markers at 4 weeks after biomaterials implantation in vivo. Red arrow, positive cell. Scale bar = 100 μm. M, material. e Semiquantification of positively stained cells in ( d ). n = 5, **** P < 0.0001. f Immunofluorescent (IF) staining of CD83 (red), CD40 (green) and nuclei (blue) after DCs seeded on the BCP or β -TCP for 24 h. Scale bar = 5 μm. g Semiquantification of positively stained cells in ( f ). n = 5, **** P < 0.0001, ns: not significant. h Relative mRNA expressions of DC maturation genes (MHC II, CD86, CD40, IL-12). n = 3, ** P < 0.01, *** P < 0.001, **** P < 0.0001. i-l FACS of T cell proliferation (Ki67 +) and activation (IFN-γ) and semiquantification of positive cell rate. n = 6, *** P < 0.001, **** P < 0.0001, ns: not significant

Article Snippet: The primary antibodies were listed as: CD11c (1:200 for IF and 1:350 for IHC, Cat: 97,585, CST, USA), F4/80 (1:250, Cat: 70,076, CST, USA), Col1a1(1:100, Cat: SA2005, Boster, China), Runx2 (1:200, Cat: ab76956, Abcam, USA), CD146 (1:100, Cat: A13927, ABclonal, USA), CD83 (1:100, Cat: A2040, ABclonal, USA), CD40 (1:100, Cat: A0218, ABclonal, USA), iNOS (1:100, Sant Cruz, USA.), Arg1 (1:150, Cat: A11925, ABclonal, USA), CD3 (1:100, Cat: PB0112, Boster, China).

Techniques: Activation Assay, In Vivo, Immunohistochemistry, Marker, Staining

Journal: Nano-Micro Letters

Article Title: Dual-Wavelength Photosensitive Nano-in-Micro Scaffold Regulates Innate and Adaptive Immune Responses for Osteogenesis

doi: 10.1007/s40820-020-00540-z

Figure Lengend Snippet: a Schematic diagram of BCP-GNCs-mediated macrophage polarization and DC maturation by releasing IL-4 and DXMS. b IF staining of Arg1 (red) and iNOS (green) of macrophage, CD83 (red) and CD40 (green) of DC and nuclei (blue) after macrophages or DCs seeded on the BCP with 690 nm far-red or 808 nm NIR stimulating the release of PBS (BCP-PBS), 690 nm far-red stimulating the release of IL-4 (BCP-IL4) or 808 nm NIR stimulating the release of DXMS (BCP-DXMS) for 24 h. Scale bar = 5 μm. c, d Semiquantification of positively stained cells in ( b ). n = 5, ** P < 0.01, *** P < 0.001. e Flowchart of time course for irradiating BCP-GNC in vivo. f–h IHC staining of M1, M2 macrophages (iNOS and Arg1), mature DCs (CD40), T cells (CD3), MSCs and osteoblasts (CD146, Runx2) under the BCP-GNCs implant in vivo. Scale bar = 100 μm. Red arrow, positive cells. M, material. i – k Semiquantification of positively stained cells in (E, F, G). n = 5, * P < 0.05, ** P < 0.01, **** P < 0.0001. l H&E and Masson staining of implant area of BCP-Con and BCP-GNCs in vivo after 4 weeks (4 W) (the red dash line shows the new bone formation area; NB, new bone; M, material). m Semiquantification of new bone area in ( k ). n = 5, **** P < 0.0001

Article Snippet: The primary antibodies were listed as: CD11c (1:200 for IF and 1:350 for IHC, Cat: 97,585, CST, USA), F4/80 (1:250, Cat: 70,076, CST, USA), Col1a1(1:100, Cat: SA2005, Boster, China), Runx2 (1:200, Cat: ab76956, Abcam, USA), CD146 (1:100, Cat: A13927, ABclonal, USA), CD83 (1:100, Cat: A2040, ABclonal, USA), CD40 (1:100, Cat: A0218, ABclonal, USA), iNOS (1:100, Sant Cruz, USA.), Arg1 (1:150, Cat: A11925, ABclonal, USA), CD3 (1:100, Cat: PB0112, Boster, China).

Techniques: Staining, In Vivo, Immunohistochemistry

Journal: Cell Transplantation

Article Title: Cyclooxygenase 2 Promotes Proliferation and Invasion in Ovarian Cancer Cells via the PGE2/NF-κB Pathway

doi: 10.1177/0963689719890597

Figure Lengend Snippet: Effects of cyclooxygenase 2 (COX2) on prostaglandin E2 (PGE2), PGE2 Receptor 2 (PTGER2) and p-nuclear factor-kappa B (NF-κB) p65 expression. (A) PGE2 release into the culture supernatants was measured by enzyme-linked immunosorbent assay. (B) The expression level of PTGER2 was estimated by quantitative polymerase chain reaction (qPCR). (C) The expression level of PTGER2, NF-κB p65, and p-NF-κB p65 and western blotting. The data are presented as the means±SDs.

Article Snippet: Membranes were blocked with 5% non-fat dry milk and were then incubated with primary antibodies specific for GAPDH (1:1000, Cell Signaling Technologies, cat. no. 2118), COX2 (1:1000, Cell Signaling Technologies, cat. no. 12282), p-NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 3031), NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 8242), CYP19 (1:1000, Santa Cruz, Dallas, TX, USA, cat. no. sc-374176), C-MYC (1:1000, Cell Signaling Technologies, cat. no. 5605), STAT3 (1:1000, Cell Signaling Technologies, cat. no. 9139), p-STAT3 (1:2000, Cell Signaling Technologies, cat. no. 9145), MMP2 (1:1000, Cell Signaling Technologies, cat. no. 4022), MMP9 (1:1000, Cell Signaling Technologies, cat. no. 3852), and PTGER2 (1:400, Bosterbio Technologies, Pleasanton, CA, USA, cat. no. BM5194) overnight at 4°C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell Transplantation

Article Title: Cyclooxygenase 2 Promotes Proliferation and Invasion in Ovarian Cancer Cells via the PGE2/NF-κB Pathway

doi: 10.1177/0963689719890597

Figure Lengend Snippet: Prostaglandin E2 Receptor 2 (PTGER2) and nuclear factor-kappa B (NF-κB) inhibitors inhibit the cyclooxygenase 2 (COX2)-promoted proliferation and invasion of ovarian cancer cells. (A) Cell viability was measured by a CCK-8 assay. a Compared Lenti-COX2 with Lenti-COX2+AH6809 and compared Lenti-COX2 with Lenti-COX2+BAY11-7802 separately, p < 0.05. (B) and (C) Representative immunohistochemical (IHC) staining of SKOV3 and OVCAR3 cells conducted with an antibody against Ki67. Magnification: ×400. (D) and (E) SKOV3 and OVCAR3 cells were seeded in 24-well Transwell chambers coated with Matrigel and cultured for 24 hours. Cell invasion was estimated. (F) The expression levels of CYP19, C-MYC, STAT3, p-STAT3, matrix metalloproteinase 2 (MMP2), and MMP9 were estimated by western blot analysis. The data are presented as the means±SDs.

Article Snippet: Membranes were blocked with 5% non-fat dry milk and were then incubated with primary antibodies specific for GAPDH (1:1000, Cell Signaling Technologies, cat. no. 2118), COX2 (1:1000, Cell Signaling Technologies, cat. no. 12282), p-NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 3031), NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 8242), CYP19 (1:1000, Santa Cruz, Dallas, TX, USA, cat. no. sc-374176), C-MYC (1:1000, Cell Signaling Technologies, cat. no. 5605), STAT3 (1:1000, Cell Signaling Technologies, cat. no. 9139), p-STAT3 (1:2000, Cell Signaling Technologies, cat. no. 9145), MMP2 (1:1000, Cell Signaling Technologies, cat. no. 4022), MMP9 (1:1000, Cell Signaling Technologies, cat. no. 3852), and PTGER2 (1:400, Bosterbio Technologies, Pleasanton, CA, USA, cat. no. BM5194) overnight at 4°C.

Techniques: CCK-8 Assay, Immunohistochemical staining, Immunohistochemistry, Cell Culture, Expressing, Western Blot