Journal: Biomolecules

Article Title: Glycogen Hydrogel Loaded with Schistosoma japonicas Peptide SJMHE1 Improves Skin Wound Healing

doi: 10.3390/biom16030392

Figure Lengend Snippet: Macrophages treated with SJMHE1 promote fibroblast and HUVEC migration through TGF-β1/Smad3 pathway and VEGFA expression. ( A ) TGF-β1 protein expression in RAW264.7 cells assessed by Western blotting. ( B ) p-Smad3 protein expression in L929cells assessed by Western blotting. ( C ) Relative mRNA expression of COL1a1 in L929 cells. ( D ) Cell scratch assay and number of migrating L929 cells (scale bar = 200 μm). ( E ) p-Smad3 protein expression in HUVECs assessed by Western blotting. ( F ) Cell scratch assay and migration of HUVECs (scale bar = 200 μm). ( G ) VEGFA expression in THP-1-induced macrophages. ( H ) Representative images of vascular network formation and tube length quantification in HUVECs in vitro (scale bar = 200 μm). Data are presented as mean ± SD ( n = 3). ns = no significance, * p < 0.05, ** p < 0.01, and *** p < 0.001. Original Western blot images are provided in the .

Article Snippet: After washing with PBS, L929 cells and HUVECs were treated with the following low-serum (2% FBS) conditioned media: CM-Control, CM-SJMHE1, CM-SJMHE1 supplemented with 5 μg/mL TGF-β1 neutralising antibody (BE0057, Bio X Cell, Lebanon, PA, USA), or normal medium supplemented with 10 ng/mL TGF-β1 recombinant protein (HY- P70648 , MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Migration, Expressing, Western Blot, Wound Healing Assay, In Vitro

Journal: Biomolecules

Article Title: Glycogen Hydrogel Loaded with Schistosoma japonicas Peptide SJMHE1 Improves Skin Wound Healing

doi: 10.3390/biom16030392

Figure Lengend Snippet: SJMHE1-gel treatment increases TGF-β1, p-Smad3, and VEGFA expression in mice. ( A ) Representative immunohistochemical staining images of TGF-β1, p-Smad3, and VEGFA on day 7 (scale bar = 50 μm). ( B ) Quantification of TGF-β1, p-Smad3 ( C ) and VEGFA ( D ) expression. Data are presented as mean ± SD ( n = 6). ns = no significance, * p < 0.05, ** p < 0.01.

Article Snippet: After washing with PBS, L929 cells and HUVECs were treated with the following low-serum (2% FBS) conditioned media: CM-Control, CM-SJMHE1, CM-SJMHE1 supplemented with 5 μg/mL TGF-β1 neutralising antibody (BE0057, Bio X Cell, Lebanon, PA, USA), or normal medium supplemented with 10 ng/mL TGF-β1 recombinant protein (HY- P70648 , MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet: Validation of co-signaling receptor modulation in independent mouse models, related to Figure 2 (A) Experimental setup. 10 6 naive Cor93 T (Cor93 T N ) cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, indicated groups of mice were injected intraperitoneally with PBS or 100 μg of monoclonal antibodies (mAbs) blocking PD-1, LAG-3, or CTLA-4. Livers were collected and analyzed at day 5. (B) Total numbers of intrahepatic leukocytes (IHLs) isolated from the indicated mice. (C) Total numbers of Cor93 T cells in the livers of the indicated mice. (D) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. Numbers represent the percentage of cells within the indicated gates. (E) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (F) Amount of serum alanine transaminases (sALTs) in the serum of the indicated groups of mice at the indicated time points. (G) Experimental setup. HBV replication-competent transgenic mice (lineage 1.3.32) were injected intraperitoneally with PBS or with 100 μg of agonist mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 4. (H) Total numbers of IHL isolated from the indicated mice. (I) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show hematoxylin-eosin (H&E) staining, the middle panels show immunohistochemical staining for cleaved caspase 3 (ΔCas3, brown), and the lower panels show immunohistochemical staining for HBcAg (brown). Scale bar represents 100 μm. (J) HBV DNA quantification by southern blot analysis of liver lysates from the indicated mice. Bands corresponding to the expected size of the integrated transgene (Tg), relaxed circular (RC), double-stranded (DS) linear, and single-stranded (SS) HBV DNAs are indicated. (K) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. (L) Experimental setup. 10 6 Cor93 T N cells were adoptively transferred into HBV-Tg mice (lineage MUP-core). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating ICOS, OX40, or 4-1BB. Livers were collected and analyzed at day 5. (M) Total numbers of IHL isolated from the indicated mice. (N) Numbers of Cor93 T cells isolated from the liver of the indicated mice. (O) Representative density plots of IFN-γ expression among Cor93 T cells in the liver of the indicated mice. (P) Total number of IFN-γ-producing Cor93 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control. (Q) Amount of sALT in the serum of the indicated group of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with control group (simple effect within row). (R) Experimental setup. 10 6 naive Env28 CD8 + TCR transgenic cells (Env28 T N ) were adoptively transferred into HBV replication-competent transgenic mice (lineage 1.3.32; background C57BL/6 × BALB/c H-2 bxd hybrids). 24 h later, selected groups of mice were injected intraperitoneally with PBS or 100 μg of mAbs activating OX40 or 4-1BB. Livers were collected and analyzed at day 5. (S) Numbers of IHL isolated from the indicated mice. (T) Numbers of Env28 T cells isolated from the liver of the indicated mice. (U) Representative density plots of IFN-γ expression among Env28 T cells in the liver of the indicated mice. (V) Percentages of IFN-γ-producing Env28 T cells in the livers of the indicated mice upon ex vivo cognate peptide stimulation. n = 3–4; one-way Brown-Forsythe and Welch ANOVA test with Dunnett correction. Each group was compared with PBS-injected controls. (W) Amount of sALT in the serum of the indicated groups of mice at the indicated time points. n = 3–4; two-way ANOVA test with Dunnett correction for multiple comparisons. Each group was compared with PBS-injected controls (simple effect within row). (X) Representative micrographs of liver sections from the indicated groups of mice. The upper panels show staining for HBcAg, and the lower panels show staining for cleaved caspase 3 (ΔCas3). Scale bar represents 100 μm. (Y and Z) Representative histograms (Y) and percentages (Z) of in vitro differentiated Cor93 T effector (Cor93 T E ) or Env28 T effector (Env28 T E ) cells producing IFN-γ upon cognate in vitro peptide stimulation at the indicated concentrations. Results are representative of two independent experiments giving similar results.

Article Snippet: InVivoMAb anti-mouse/human/rat/monkey ICOS (CD278) , Bio X Cell , Cat# BE0353; RRID: AB_2894772.

Techniques: Biomarker Discovery, Injection, Bioprocessing, Blocking Assay, Isolation, Expressing, Ex Vivo, Transgenic Assay, Staining, Immunohistochemical staining, Southern Blot, Control, In Vitro

Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet:

Article Snippet: InVivoMAb anti-mouse/human/rat/monkey ICOS (CD278) , Bio X Cell , Cat# BE0353; RRID: AB_2894772.

Techniques: Purification, Virus, Recombinant, Staining, Saline, Fluorsave, Sequencing, DNA Labeling, Cell Isolation, Sample Prep, Reverse Transcription, Software, Microscopy

Journal: Inflammation and Regeneration

Article Title: Evaluation of immunosuppression protocols for MHC-matched allogeneic iPS cell-based transplantation using a mouse skin transplantation model

doi: 10.1186/s41232-021-00190-7

Figure Lengend Snippet: Blockade of leukocyte costimulatory molecules permits long-term engraftment of mouse skin grafts from MHC-matched allogeneic donors. ( A ) CB-based immunoregulation protocol. Immunosuppression was performed by using a protocol of administrating a combination of anti-CD40L mAb and CTLA4-Ig daily (CB) with or without rapamycin on every 3 days from POD 1 (CB + rapa). ( B ) Skin graft survival under immunosuppression with CB ( n = 9, C3129F1; n = 9, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( C ) Skin graft survival under immunosuppression with CB + rapa ( n = 9, C3129F1; n = 8, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( D ) Macroscopic observation (upper panels) and hematoxylin and eosin stained section (lower panels) of autologous (C3129F1), CBA/N, and C57BL/6 skin grafts on day 100. Scale bars: 100 μm. ( E ) Immunohistochemical staining for CD3 of C57BL/6 skin grafts harvested from CB or CB + rapa treated groups on 100 days post-transplantation. Upper and lower panels represent CD3 specific staining and isotype control stained section, respectively. Scale bars: 100 μm. ( F ) Recipient T cell response in MHC-matched but minor antigen-mismatched skin transplantation. The T cell proliferation rates in each treatment group were normalized to that of C3129F1 mice stimulated with autologous irradiated splenocytes. Error bars indicate standard error of technical triplicates. Similar results were obtained in two independent experiments. * p < 0.05, ** p < 0.01 (Tukey’s HSD test). ( G ) De novo anti-donor antibody production in the recipients. Error bars indicate standard error of biological replicates ( n = 8, naive; n = 4, non-treatment; n = 9, CB; n = 9, CB + rapa). tx, transplantation; mAb, monoclonal antibody; CB, co-stimulatory molecule blocking; rapa, rapamycin; POD, post-operative day

Article Snippet: For co-stimulatory molecule blockade (CB) therapy, anti-CD40 ligand (CD40L) (MR-1 for mouse and #BE0292 for human, BioXcell) and CTLA4Ig (Orencia®, Bristol Myers Squibb for mouse and #BE0099, BioXcell for human) were administered at a dose of 500 μg for anti-CD40L, 400 μg for CTLA4Ig on days 0, 2, 4, and 6 after transplantation.

Techniques: Staining, Immunohistochemical staining, Transplantation Assay, Control, Irradiation, Blocking Assay