Journal: Cell reports

Article Title: Integrating Gene and Protein Expression Reveals Perturbed Functional Networks in Alzheimer’s Disease

doi: 10.1016/j.celrep.2019.06.073

Figure Lengend Snippet:

Article Snippet: Normal Horse Serum , Vector Laboratories , Cat# S-2000; RRID: AB_2336617.

Techniques: Plasmid Preparation, Recombinant, Software, Microscopy

Journal: Cell

Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

doi: 10.1016/j.cell.2018.08.067

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Article Snippet: Slides were washed in PBS containing 0.05% Tween 20 (PBS-T) and incubated with ImmPRESS HRP Anti-Rabbit or Anti-Mouse IgG (Peroxidase) Polymer Detection Kit (Vector Laboratories, Peterborough, U.K) for thirty minutes at room temperature.

Techniques: Recombinant, Coagulation, Antibody Labeling, Plasmid Preparation, Blocking Assay, Staining, Imaging, Software, Hybridization

Journal: Plant Physiology

Article Title: ABI5-BINDING PROTEIN2 Coordinates CONSTANS to Delay Flowering by Recruiting the Transcriptional Corepressor TPR2 ¹

doi: 10.1104/pp.18.00865

Figure Lengend Snippet: AFP2 reinforces CO and TPR2 to coordinate flowering time. A, Y3H analysis to detect the formation of the CO-AFP2-TPR2 complex. Yeast cotransformed with these three constructs could grow well on the nonselective medium lacking Leu, Trp, and uracil (−L/−W/−U), but only yeast harboring constructs that had positive interactions were able to grow on restrictive growth medium supplemented with 10 mm 3-aminotriazole plus 2% (w/v) Gal and lacking His/Leu/Trp/Ura (−H/−L/−W/−U). B, In vitro pull-down analysis of the interactions among CO, AFP2, and TPR2. Recombinant GST-TPR2 and MBP-CO proteins were produced in E. coli. After cell lysis, cell extracts of GST-TPR2 and MBP-CO were mixed with HIS-AFP2, HIS-AFP∆E, or HIS-AFP2∆J, respectively, and then incubated with magnetic anti-His-coupled magnetic beads. His-tagged full-length or truncated AFP2 was precipitated and washed using a magnetic stand, eluted by boiling in SDS loading buffer, and separated by SDS-PAGE. GST-TPR2 and MBP-CO were detected by immunoblotting. C, Co-IP analysis of the CO-AFP2-TPR2 complex in vivo. The CO-HA/afp2 transgenic line was crossed with AFP-ox/afp2 to obtain CO-HA/AFP2-ox/afp2, which was subjected to Co-IP analysis. Total proteins were extracted from CO-HA/AFP2-ox/afp2 and immunoprecipitated with anti-Flag agarose beads, and the immunoprecipitated proteins were detected with anti-TPR2 antibody. D, Flowering phenotype of the afp2 mutant and the indicated transgenic lines in the afp2 mutant background. The photos were taken at 18 d after seeds germination. Bar = 3 cm. E, Flowering times based on the total rosette leaf number under LD conditions. Data are means ± sd of three biological replicates. For each line, 20 plants were observed. Bars with different letters are significantly different at P < 0.05 (Tukey’s test). F, RT-qPCR analysis of CO transcript levels in the afp2 mutant and different transgenic lines in the afp2 background. IPP2 was used as an internal control. Data are means ± sd of three biological replicates. Bars with different letters are significantly different at P < 0.05 (Tukey’s test). 3AT, 3-aminotriazole.

Article Snippet: The pulled-down proteins were extensively washed with buffer (50 m m Tris-HCl, pH 7.4, 100 m m NaCl, and 0.6% [v/v] Triton X-100) before the samples were resolved on 8% (w/v) SDS-PAGE gels and analyzed by protein gel blot analysis using anti-MBP (1:5000, New England Biolabs), anti-HIS (1:300, Qiagen), and anti-GST (1:3000, Invitrogen), followed by a mouse secondary antibody (1:5000, Promega) and the ECL system (Invitrogen).

Techniques: Construct, In Vitro, Recombinant, Produced, Lysis, Incubation, Magnetic Beads, SDS Page, Western Blot, Co-Immunoprecipitation Assay, In Vivo, Transgenic Assay, Immunoprecipitation, Mutagenesis, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Plasmid Preparation, Construct, SDS Page, Molecular Weight, Recombinant, Flow Cytometry, Expressing, In Vitro, Cell Culture

Journal: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: a. Frequencies of immune cell subsets. b. SCENIC prediction of IRF4 and Blimp1 transcription factor activity. c. Immunoglobulin gene expression. d. UMAP plots showing cell cycle gene signature scores. e. Quantification of cell cycle gene signature scores. f. Scaled signature scores for TGF-β–inducible genes, senescence genes, and apoptosis-related genes, aggregated by sample.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Activity Assay, Gene Expression