Journal: iScience

Article Title: Uhrf1 governs the proliferation and differentiation of muscle satellite cells

doi: 10.1016/j.isci.2022.103928

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Myh3 , DSHB , cat# F1.652; RRID: AB_528358.

Techniques: Recombinant, Western Blot, Imaging, Multiplex Assay, Library Quantification, Methylation, Electrophoresis, Software

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 1 Expression of CD30 in the T cells and B cells and generation of CD30-deficient mice. (A) The indicated hematopoietic cells were sorted from wild-type mice. The level of CD30 mRNA expression was measured by qRT-PCR, using β-Actin as internal control. (B) The frequency of CD30+ T cells in CD4 T cells upon TCR stimulation (left). The overlays show CD30 surface expression of CD4 T cells upon TCR stimulation (right). (C) The frequency of CD30+ T cells in CD8 T cells upon TCR stimulation (left). The overlays show CD30 surface expression of CD8 T upon TCR stimulation (right). (D) CD30 expression level was detected by qRT-PCR. (E) Western blotting analysis of CD30 expression in splenic T cells. Purified splenic T cells were stimulated for 48 h on tissue culture plate coated with anti-CD3 (10 μg/mL) plus anti-CD28 (5 μg/mL) and IL-2 (100 ng/mL). The cell lysates were subjected to direct Western blot analysis with the indicated antibodies. Data shown are representative of or obtained from 3 (A–E) independent experiments. Mean ± SD is shown.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot, Purification

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 3 Serum immunoglobulin levels and serum antibody response to TD antigens in CD30-KO and wild-type mice. (A) Serum immunoglobulin isotype levels in wild-type and CD30-KO mice were measured by ELISA. (B) Serum NP-specific immunoglobulin isotype response to the TD antigen NP-KLH immunization in wild-type and CD30-KO mice. Sera were collected at the indicated time points after immunization and antigen-specific antibodies were measured by ELISA. (C) CD30-KO and wild-type mice immunized with NP-KLH. Splenocytes from CD30-KO and wild-type mice were stained with anti-B220, anti-CD95, and anti-GL7 at day 9 after immunization and analyzed by FACS. Percentages indicate cells in the gated B220+ cells. (D) Dot plots show percentages and numbers of GC B cells in the spleen of CD30-KO and wild-type mice. (E) Splenic mature B cells were stimulated with LPS for 48 h, and cell lysates were subjected to direct Western blot analysis with the indicated antibodies. (F) The intensities of protein bands were quantitated using Image-J software. Data shown are representative of or obtained from 6 (A), 4 (B, E, F), 5 (C, D), or 4 (E, F) CD30-KO and wild-type littermates. Mean ± SD is shown. *p < .05, **p < .01.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Software

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 4 CD30 deficiency has no effect on T-cell development. (A) Dot plots show numbers of total thymocytes in CD30-KO and wild- type mice. (B) Thymocytes from CD30-KO and wild-type mice were stained with anti-CD4 and anti-CD8 and analyzed by FACS. Percentages indicate cells in the gated live cells. (C) Dot plots show percentages and numbers of DN, DP, CD4, and CD8 T cells in the thymocytes of CD30-KO and wild-type mice. (D) Thymocytes from CD30-KO and wild-type mice were stained with anti-CD4, anti-CD8, anti-CD25, and anti-CD44 and analyzed by FACS. Percentages indicate cells in the gated DN T cells. (E) Dot plots show percentages and numbers of DN1, DN2, DN3, and DN4 T cells in the DN T cells of CD30-KO and wild-type mice. (F) Splenocytes from CD30-KO and wild-type mice were stained with anti-CD4 and anti-CD8 and analyzed by FACS. Percentages indicate cells in the gated live cells. (G) Dot plots show percentages and numbers of CD4 and CD8 T cells in the spleen of CD30-KO and wild-type mice. Data shown are representative of or obtained from 10 (A–G) CD30-KO and wild-type littermates. Mean ± SD is shown.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Staining

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 5 CD30 deficiency reduces TCR-mediated proliferation. (A) Splenic T cells were labeled with CFSE, according to the manufacturer's recommendation, and stimulated as indicated for 72 h before analysis for cell proliferation (left). Dot plots show the percentages of proliferate rate (right). (B) Splenic T cells were stimulated as indicated for 72 h, stained with PI, and analyzed for cell cycle profile by FACS (left). Dot plots show percentages of cells in S/G2/M phase after stimulation (right). Data shown are representative of or obtained from 3 (A, B) independent experiments. Mean ± SD is shown. *p < .05, ** p < .01.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Labeling, Staining

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 6 CD30 deficiency reduces TCR-mediated T-cell activation. (A) Total thymocytes were stimulated with anti-CD3 (10 μg/ mL) for the indicated time and cell lysates were subjected to direct Western blot analysis with the indicated antibodies. (B) The intensities of protein bands were quantitated using Image-J software. (C, D) Splenic T cells from CD30-KO and wild-type littermates were stimulated with anti-CD3 for 72 h and analyzed by FACS and ELISA. (C) Splenic T cells were stained with anti-CD4, anti-CD8, anti-CD25, and anti- CD69 upon TCR stimulation. Dot plots show percentages of cells in CD4 and CD8 T cells after stimulation. (D) ELISA analysis of IL-6 and IFN-γ released in the splenic T cell culture supernatant. Data shown are representative of or obtained from 6 (A, B) or 3 (C, D) independent experiments. Mean ± SD is shown. *p < .05.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Activation Assay, Western Blot, Software, Enzyme-linked Immunosorbent Assay, Staining, Cell Culture

Journal: The FASEB Journal

Article Title: CD30 plays a role in T‐dependent immune response and T cell proliferation

doi: 10.1096/fj.202301747rr

Figure Lengend Snippet: FIGURE 7 Analysis of CD30-KO gene expression signature in TCR-mediated T cells. (A, B) Splenic T cells from CD30-KO and wild- type littermates were sorted and stimulated with anti-CD3/anti-CD28/IL-2 for 48 h and subjected to high-throughput RNA-sequencing. (A) Volcano plot of differentially expressed genes between CD30-KO and wild-type T cells. (B) Significantly downregulation pathways from the KEGG pathway database in CD30-KO and wild-type T cells. (C) The relative mRNA levels of LC3 and Atg7 were quantified by qRT-PCR in CD30-KO and wild-type T cells with anti-CD3/anti-CD28/IL-2 stimulation. The relative mRNA levels of the indicated genes in CD30-KO and wild-type T cells were normalized to β-Actin. (D) Splenic T cells were stimulation with anti-CD3/anti-CD28/IL-2 and cell lysates were subjected to direct Western blot analysis with the indicated antibodies (left). The intensities of protein bands were quantitated using Image-J software (right). Data shown are representative of or obtained from 2 (A, B) or 3 (C, D) independent experiments. Mean ± SD is shown. *p < .05, **p < .01.

Article Snippet: Rabbit monoclonal anti- CD30 (#54535) was purchased from Cell Signaling Technology.

Techniques: Gene Expression, High Throughput Screening Assay, RNA Sequencing, Quantitative RT-PCR, Western Blot, Software