Journal: ACS Omega

Article Title: Proteomics-Based Studies Identify the Metallothionein-2A as a Potential Target of Thioxo-Tetrahydropyrimidine Derivatives in Hepatocellular Carcinoma

doi: 10.1021/acsomega.5c05138

Figure Lengend Snippet: A, Molecular structure of THPP and DHPM compounds. B, Cytotoxicity effect of individual compounds in 10 cancer cell lines including brain cancer (SW-1088 and U87 MG), breast cancer (MDA-MB-231 and MCF7), lung cancer (H460 and A-549), liver cancer (HepG2 and HuH-7), and colon cancer (HT-29 and HCT 116). All cancer cell lines were tested with various doses of compounds (0–100 μM) for 24 h. Cell viability was evaluated by MTT assay (mean ± SD).

Article Snippet: Ten cancer cell lines including SW-1088 (lot no. 70036445), U87 MG (lot no. 70029548), H460 (lot no. 70039818), A-549 (lot no. 70035208), MCF7 (lot no. 70033778), MDA-MB-231 (lot no. 70029549), HepG2 (lot no. 70039681), HT-29 (lot no. 70019050), and HCT 116 (lot no. 70040763) were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: MTT Assay

Journal: ACS Omega

Article Title: Proteomics-Based Studies Identify the Metallothionein-2A as a Potential Target of Thioxo-Tetrahydropyrimidine Derivatives in Hepatocellular Carcinoma

doi: 10.1021/acsomega.5c05138

Figure Lengend Snippet: Effect of compounds 3d and 4d on apoptosis in liver cancer cell lines. HepG2 cells were treated with 0, 10, 20, and 40 μM, and HuH-7 cells were treated with 0, 5, 10, and 20 μM for 24 h. A., Phase contrast and Hoechst 33342/PI staining in HepG2 and HuH-7 cells after treatment with 3d and 4d were detected by fluorescence microscopy (scale bar = 100 μm). B., Apoptotic cells of HepG2 and HuH-7 after treatment with 3d and 4d at their IC 50 concentrations were assessed by Annexin V/PI flow cytometry. C., Western blot analysis of PARP and cleaved PARP expression in both cell lines normalized to β-actin. D, E. Colony formation of HepG2 and HuH-7 after treatment with 3d and 4d for 24 h with concentrations of 10 and 20 μM for HepG2 cells and 5 and 10 μM for HuH-7 cells. */# = p < 0.05, **/## = p < 0.01, ***/### = p < 0.001.

Article Snippet: Ten cancer cell lines including SW-1088 (lot no. 70036445), U87 MG (lot no. 70029548), H460 (lot no. 70039818), A-549 (lot no. 70035208), MCF7 (lot no. 70033778), MDA-MB-231 (lot no. 70029549), HepG2 (lot no. 70039681), HT-29 (lot no. 70019050), and HCT 116 (lot no. 70040763) were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Staining, Fluorescence, Microscopy, Flow Cytometry, Western Blot, Expressing

Journal: ACS Omega

Article Title: Proteomics-Based Studies Identify the Metallothionein-2A as a Potential Target of Thioxo-Tetrahydropyrimidine Derivatives in Hepatocellular Carcinoma

doi: 10.1021/acsomega.5c05138

Figure Lengend Snippet: Proteomic analysis of liver cancer cells treated with compounds 3d and 4d at concentrations of 20 μM for 12 h. A,-B. Heatmaps showing hierarchical clustering of differentially expressed proteins in HepG2 (A) and HuH-7 (B) cells treated with 3d, 4d, or control. C–,D. Principal component analysis (PCA) of proteomic profiles in HepG2 (C) and HuH-7 (D) cells. E–-H. Canonical pathway enrichment analysis using Ingenuity Pathway Analysis (IPA) for HepG2 (E) and HuH-7 (F) treated with 3d, and HepG2 (G) and HuH-7 (H) treated with 4d. Bubble color indicates activation z-score (orange = activated; blue = inhibited), and the size corresponds to the number of overlapping proteins.

Article Snippet: Ten cancer cell lines including SW-1088 (lot no. 70036445), U87 MG (lot no. 70029548), H460 (lot no. 70039818), A-549 (lot no. 70035208), MCF7 (lot no. 70033778), MDA-MB-231 (lot no. 70029549), HepG2 (lot no. 70039681), HT-29 (lot no. 70019050), and HCT 116 (lot no. 70040763) were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Control, Activation Assay

Journal: ACS Omega

Article Title: Proteomics-Based Studies Identify the Metallothionein-2A as a Potential Target of Thioxo-Tetrahydropyrimidine Derivatives in Hepatocellular Carcinoma

doi: 10.1021/acsomega.5c05138

Figure Lengend Snippet: Identification of MT-2A as a potential molecular target of compounds 3d and 4d. A, Venn diagram illustrating the overlap of differentially expressed proteins (DEPs) among HepG2 and HuH-7 cells treated with compounds 3d and 4d at a concentration of 20 μM for 12 h. The bar plots below show the number of unique and shared DEPs across conditions. B–-C., Bar graphs displaying the log 2 fold change of the top upregulated and downregulated DEPs from the overlapping set of DEPs between 3d and 4d treatments in HepG2 (B) and HuH-7 (C) cells. Notably, MT-2A was among the most highly upregulated overlapping proteins in both cell lines. D-–F. The binding orientations of the (R)-isomers of compounds 3d (D), 4d (E), and 19 (F) with metallothionein-2. Amino acid residues that showed hydrogen bonding as green dotted lines are depicted as sticks. The amino acids are labeled in red. The Discovery Studio 4.5 visualization software was used to generate the graphical contents.

Article Snippet: Ten cancer cell lines including SW-1088 (lot no. 70036445), U87 MG (lot no. 70029548), H460 (lot no. 70039818), A-549 (lot no. 70035208), MCF7 (lot no. 70033778), MDA-MB-231 (lot no. 70029549), HepG2 (lot no. 70039681), HT-29 (lot no. 70019050), and HCT 116 (lot no. 70040763) were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Concentration Assay, Binding Assay, Labeling, Software

Journal: bioRxiv

Article Title: A surrogate marker of protection confirms the efficacy of an AddaS03-adjuvanted West Nile virus subunit vaccine

doi: 10.64898/2026.04.20.719748

Figure Lengend Snippet: ( A ) Identification of purified sEnv by CBB staining and immunoblotting using a pan-orthoflavivirus anti-fusion loop monoclonal antibody (clone 4G2). ( B ) Five-week-old BALB/c mice (5-week-old, n=6) were immunized subcutaneously with a vaccine formulation containing various antigen doses (0, 0.1, 0.5, 1, or 5 μg) with or without adjuvant (Alhydrogel or AddaS03) twice at 2 weeks interval. Serum samples were collected at 13 and 35 days after the first immunization. ( C to F ) Binding and neutralizing antibody titers in murine sera after priming ( C and E ) and boosting immunizations ( D and F ) were determined by indirect ELISA using WNV SVP and by a neutralization assay, respectively. Dotted line indicates the LOD. The threshold titer estimated as sufficient to protect against lethal WNV infection in is also indicated as the LOD, since the threshold value was below the LOD. Data are represented as the mean±SD of biological replicates (n=6). Statistical analysis was performed using two-way ANOVA followed by Dunnett’s multiple comparison test. All statistical differences are determined in comparison with the sEnv alone (gray) or Alhydrogel (blue) group within the same antigen dose (* p <0.01, ** p <0.005, **** p <0.0005). ns, not significant.

Article Snippet: The membranes were blocked with 5% skimmed milk in PBST and incubated with pan-orthoflavivirus anti-fusion loop monoclonal antibody 4G2 (supernatant from D1-4G2-4-15 cells, ATCC) at 4°C overnight.

Techniques: Purification, Staining, Western Blot, Formulation, Adjuvant, Binding Assay, Indirect ELISA, Neutralization, Infection, Comparison

Journal: Nature Communications

Article Title: Immune-induced TCR-like antibodies regulate specific T cell response in mice

doi: 10.1038/s41467-026-71384-1

Figure Lengend Snippet: a Production of iTabs by immunization with mutated FR – or FR + HEL peptides ( n = 4 mice per group). b Binding of serum Abs from mice immunized with mutated FR – or FR + HEL peptides against WT LK35.2(black line), H2-DMα (blue line) or invariant chain (red line) knockout cells pulsed with HEL protein. c Blocking of 3A9 TCR-Fc binding by Abs induced by mutated FR – or FR + HEL peptide immunization. d Inhibition of HEL-specific T cell response by immunization with mutated FR + HEL peptide. Mice immunized with mutated FR – or FR + HEL peptide were further immunized with WT HEL peptides. IL-2 production (left) and T cell proliferation (right) upon FR + HEL peptide stimulation were shown ( b , c and d , n = 3 technical replicates). p- values were determined by two-way ANOVA for [c and d] with Tukey’s correction, except for ( a ) where a two-sided Student’s t test was used. Bars represent means (min–max) ( a ) and mean values +/− SD ( c , d ). The data represent three independent experiments.

Article Snippet: The human embryonic kidney 293 T cells (RCB2202) were purchased from the RIKEN Cell Bank, and LK35.2 cells (ATCC HB-98) expressing I-A k and NK92 cells (ATCC CRL-2407) were obtained from the American Type Culture Collection.

Techniques: Binding Assay, Knock-Out, Blocking Assay, Inhibition