Journal: Cartilage

Article Title: Semaphorin 7A Regulates the Balance Between Cartilaginous and Fibrous Tissues in the Repair Process of Articular Cartilage Damage

doi: 10.1177/19476035261418126

Figure Lengend Snippet: Integrin β1 (ITGB1) transduces Sema7A signal in chondrocytes. (A) Feature plot showing the expression distribution of PLXNC1 and ITGB1 in the UMAP plot of Figure 1 . (B) P2 Sema7a +/+ and Sema7a -/- chondrocytes were cultured in the presence or absence of neutralizing anti-ITGB1 antibody for 48 hours. Relative mRNA expression of chondrocyte-maker genes was analyzed. Error bars denote mean ± standard error. * P < 0.05, two-tailed Welch’s t test ( n = 6).

Article Snippet: Anti-integrin β1 neutralizing antibody solution (4.3 mg/ml, cat #BE0232, BioXCell, Lebanon, NH) was purchased.

Techniques: Expressing, Cell Culture, Two Tailed Test

Journal: The FASEB Journal

Article Title: Nanoliposome C6‐Ceramide in combination with anti‐CTLA4 antibody improves anti‐tumor immunity in hepatocellular cancer

doi: 10.1096/fj.202101707r

Figure Lengend Snippet: FIGURE 2 LipC6 in combination with αCTLA4 Ab suppressed HCC tumor progression. Tumor-bearing mice were randomly grouped to receive treatments with LipC6, αCTLA4 Ab, or both. Ghost (LipC6 vehicle control) and isotype Ab (αCTLA4 Ab control) were used in control group (W/O). MRI was used to detect tumor growth in the mice three months after oncogenic hepatocyte inoculation. (A) Design of treatment with LipC6, αCTLA4 Ab (αCTLA4), and their combination. (B) Representative MRI showing HCC tumor progression in response to indicated treatments. Tumor nodules are outlined by red circles. (C) Accumulated data of HCC tumor volume measured by MRI. The tumor volume was calculated by ImageJ. n = 6, *p < .05, error bars represent means ± SD. (D) Macroscopic image of tumors. Yellow arrows point to the tumors. (E) Accumulated tumor weights in four groups of mice, n = 6, **p < .01, error bars represent means ± SD. Representative images of IHC staining for cleaved caspase 3 (F) and cleaved PARP (G). Red arrows point to positive signals. Bars: 50 µm

Article Snippet: αPD- 1 Ab (CAT# BE0146, BioXCell, USA), αCTLA4 Ab (CAT# BE0131, BioXCell, USA), or their IgG isotypes (CAT# BE0089, CAT# BE0087, BioXCell, USA) were intraperitoneally injected into mice every 3 days for two weeks at 100 μg/mouse in 200 μl of PBS buffer.

Techniques: Control, Immunohistochemistry

Journal: The FASEB Journal

Article Title: Nanoliposome C6‐Ceramide in combination with anti‐CTLA4 antibody improves anti‐tumor immunity in hepatocellular cancer

doi: 10.1096/fj.202101707r

Figure Lengend Snippet: FIGURE 3 LipC6 in combination with αCTLA4 Ab activated intrahepatic effector CD8+ T cells. Tumor-bearing mice were randomly grouped to receive the indicated treatments as shown in Figure 2. Five weeks post-treatment, the mice were sacrificed, and TILs were isolated for flow cytometry. (A) Representative and accumulated frequency of tumor-infiltrating CD8+ T cells in four groups of mice. n = 3, *p < .05, **p < .01, ***p < .001, error bars represent means ± SD. (B) Representative images of IHC staining for CD8+ T cells. Red arrows point to the positive staining cells, bars: 50 µm. (C) Representative and accumulated frequency of tumor-infiltrating CD69+CD8+ T cells in four groups of mice. n = 3, *p < .05, **p < .01, ***p < .001, error bars represent means ± SD. (D) Representative and accumulated frequency of IFN-γ-producing CD8+ T cell in the TILs isolated from four groups of mice in response to TSA stimulation. The cultured TILs were stimulated with large T antigen (TAg) epitopes I and IV in the presence of Brefeldin A. Five hours post-stimulation, IFN-γ-producing CD8+ T cells were evaluated by flow cytometry. n = 3, *p < .05, error bars represent means ± SD

Article Snippet: αPD- 1 Ab (CAT# BE0146, BioXCell, USA), αCTLA4 Ab (CAT# BE0131, BioXCell, USA), or their IgG isotypes (CAT# BE0089, CAT# BE0087, BioXCell, USA) were intraperitoneally injected into mice every 3 days for two weeks at 100 μg/mouse in 200 μl of PBS buffer.

Techniques: Isolation, Flow Cytometry, Immunohistochemistry, Staining, Cell Culture

Journal: The FASEB Journal

Article Title: Nanoliposome C6‐Ceramide in combination with anti‐CTLA4 antibody improves anti‐tumor immunity in hepatocellular cancer

doi: 10.1096/fj.202101707r

Figure Lengend Snippet: FIGURE 4 LipC6 in combination with αCTLA4 Ab suppressed KLF2 expression in tumor-infiltrating CD8+ T cells. Tumor-bearing mice were randomly grouped to receive the indicated treatments as shown in Figure 2. Five weeks after the last treatment, the TILs were isolated. KLF2 expression in CD8+ T cells was detected by flow cytometry. (A) Representative histogram showing KLF2 expression in tumor- infiltrating CD8+ T cells in four groups of mice. Liver-infiltrating CD8+ T cells from normal mice were used for control. (B) Accumulated data of KLF2 expression in tumor-infiltrating CD8+ T cells in four groups of mice. Liver-infiltrating CD8+ T cells from normal mice were used for control. n = 3, ***p < .001, error bars represent means ± SD

Article Snippet: αPD- 1 Ab (CAT# BE0146, BioXCell, USA), αCTLA4 Ab (CAT# BE0131, BioXCell, USA), or their IgG isotypes (CAT# BE0089, CAT# BE0087, BioXCell, USA) were intraperitoneally injected into mice every 3 days for two weeks at 100 μg/mouse in 200 μl of PBS buffer.

Techniques: Expressing, Isolation, Flow Cytometry, Control

Journal: The FASEB Journal

Article Title: Nanoliposome C6‐Ceramide in combination with anti‐CTLA4 antibody improves anti‐tumor immunity in hepatocellular cancer

doi: 10.1096/fj.202101707r

Figure Lengend Snippet: FIGURE 5 LipC6 reduced the generation of FoxP3+ Tregs and the expression of CTLA4 and KLF2 in CD8+ T cells in vitro. Naive C57BL/6 mice were used to prepare RBC-depleted splenocytes. These cells received the indicated treatments with LipC6, αCTLA4, or both in the presence of anti-CD3 & anti-CD28 Abs for 24 h for flow cytometric assay. Ghost (LipC6 vehicle control) and isotype Ab (αCTLA4 Ab control) were used in control group (W/O). (A) Representative flow cytometry and accumulated data showing the frequency of CD8+CD3+ T cells in the splenocytes in response to the different treatments. n = 3, *p < .05, **p < .01, ***p < .001, error bars represent means ± SD. (B) Representative flow cytometry and accumulated data showing frequency of CTLA4+CD8+ T cells in the splenocytes in response to the different treatments. n = 3, *p < .05, **p < .01, error bars represent means ± SD. (C) Representative flow cytometry and accumulated data showing the frequency of CD4+CD25+ FoxP3+ Treg cells in the splenocytes in response to the different treatments. n = 3, *p < .05, **p < .01, error bars represent means ± SD. (D) Representative flow cytometry and accumulated data showing KLF2 expression in CD3+ T cells in the splenocytes in response to the different treatments. n = 3, **p < .01, error bars represent means ± SD

Article Snippet: αPD- 1 Ab (CAT# BE0146, BioXCell, USA), αCTLA4 Ab (CAT# BE0131, BioXCell, USA), or their IgG isotypes (CAT# BE0089, CAT# BE0087, BioXCell, USA) were intraperitoneally injected into mice every 3 days for two weeks at 100 μg/mouse in 200 μl of PBS buffer.

Techniques: Expressing, In Vitro, Flow Cytometry, Control

Journal: The FASEB Journal

Article Title: Nanoliposome C6‐Ceramide in combination with anti‐CTLA4 antibody improves anti‐tumor immunity in hepatocellular cancer

doi: 10.1096/fj.202101707r

Figure Lengend Snippet: FIGURE 7 Molecular actions of LipC6, αCTLA4 Ab, or their combination on T cells. Pan T cells purified from splenocytes in wild-type mice with negative selection beads. These cells respectively received the treatments of LipC6, αCTLA4 Ab or both in the presence of anti- CD3 & anti-CD28 Abs. Ghost (LipC6 vehicle control) and isotype Ab (αCTLA4 Ab control) were used in control group (W/O). 24 h later, the cells were harvested to extract total RNAs for detecting the expression of the indicated genes by qPCR. The expression level of the indicated genes was normalized to housekeeping genes. The fold changes of each gene were shown for Klf2 (A), Ctla4 (B), Pdcd1 (C), Foxp3 (D), Gzmb (E), Tnf (F), and Ifng (G). *p < .05, **p < .01, ***p < .001. n = 3, Error bars represent means ± SD. (G) Schematic diagram depicting the signal pathways mediating combinatorial function of LipC6 and αCTLA4 Ab

Article Snippet: αPD- 1 Ab (CAT# BE0146, BioXCell, USA), αCTLA4 Ab (CAT# BE0131, BioXCell, USA), or their IgG isotypes (CAT# BE0089, CAT# BE0087, BioXCell, USA) were intraperitoneally injected into mice every 3 days for two weeks at 100 μg/mouse in 200 μl of PBS buffer.

Techniques: Purification, Selection, Control, Expressing