il 12  (MedChemExpress)

Journal: Frontiers in Neurology

Article Title: Elevated interleukin-12B is associated with increased seizure susceptibility: insights from two-sample Mendelian randomization and in vivo experiment

doi: 10.3389/fneur.2025.1706857

Figure Lengend Snippet: Administration of IL-12 beta protein increases seizure susceptibility. (A) Seizure susceptibility measured by 6-Hz seizure threshold test ( n = 12 mice/group). (B) Seizure score within 30 min after picrotoxin administration ( n = 6 mice/group). (C) A representative diagram illustrating EEG, EMG, baseline, and seizure patterns after pilocarpine administration. (D) A time-frequency spectrogram of EEG corresponding to the EEG in (C) ( n = 10 mice/group). (E) Latency to SE after pilocarpine administration. (F) Survival probability within 60 min post pilocarpine administration ( n = 10 mice/group). * p < 0.05, ** p < 0.01 by Student’s t test ( A and E ), generalized linear models with Bonferroni test (B) , Kaplan–Meier analysis with Log Rank (Mantel-Cox) test (F) . Values are means ± SD ( A and E ), medians (IQR) (B) , rate (F) .

Article Snippet: Mice were randomly assigned into two groups. (1) Control group: mice received an intraperitoneal (i.p.) injected of saline (10 mL/kg); (2) IL-12B group: mice were administered an injection of IL-12 beta protein (mouse, HEK293, His; 1 μg/mouse/day, i.p., MedChemExpress, USA) for three consecutive days.

Techniques:

Journal: Frontiers in Neurology

Article Title: Elevated interleukin-12B is associated with increased seizure susceptibility: insights from two-sample Mendelian randomization and in vivo experiment

doi: 10.3389/fneur.2025.1706857

Figure Lengend Snippet: Comprehensive results of the MR analysis. (A) Causal estimates presented as odds ratios (ORs) and 95% confidence intervals for the effect of ILs on GE. (B) Scatter plots depicting the causal relationship between IL-12B and GE. (C) Detailed forest plots displaying the estimated MR effect size of IL-12B on GE. (D) Leave-one-out analysis assessing the effect of IL-12B on GE. (E) Funnel plot evaluating the potential heterogeneity between IL-12B and GE.

Article Snippet: Mice were randomly assigned into two groups. (1) Control group: mice received an intraperitoneal (i.p.) injected of saline (10 mL/kg); (2) IL-12B group: mice were administered an injection of IL-12 beta protein (mouse, HEK293, His; 1 μg/mouse/day, i.p., MedChemExpress, USA) for three consecutive days.

Techniques:

Journal: Scientific Reports

Article Title: Bromodomain and extra-terminal protein inhibitors modulate natural killer cell function and differentiation

doi: 10.1038/s41598-025-33437-1

Figure Lengend Snippet: Small-molecule drug screen identified key epigenetic regulators of NK cell function. ( A ) Overview of strategy to screen the effects of 158 small-molecule drugs targeting epigenetic regulators on secreted cytokine and cytolytic protein levels after NK-92 cell activation with IL-12 and IL-15. ( B ) Heatmap of concentrations of cytokines and cytolytic proteins in cell culture supernatants, displayed as normalized z-scores at 25µM drug treatment concentration. K-means clustering utilized to group the drugs by protein pattern into four clusters (c1-c4). ( C ) Drug names and type that induced both ≤ 0.5 fold change in analyte levels and did not reduce cell viability ≤ 80%. ( D ) Heatmap of concentrations of cytokines and cytolytic proteins in cell culture supernatants, displayed as normalized z-scores at 100nM drug concentration treatment. ( E ) Drug names and type that mediated ≤ 0.5 fold change in either IFNγ or IL-6 levels along with cell viability. Arrows highlight drugs that did not reduce cell viability ≤ 80%. All five selected drugs targeted BET proteins.

Article Snippet: Cells were stimulated with 10ng/ml recombinant IL-12 (130-129-720, Miltenyi) plus 100ng/ml recombinant IL-18 (B001-5, R&D Systems) and with 100nM CPI-203 (15479, Cayman Chemical) or 100nM AZD 5153 (15479, Cayman Chemical), or cultured in medium only for 48 h at 37 °C.

Techniques: Cell Function Assay, Activation Assay, Cell Culture, Concentration Assay

Journal: Scientific Reports

Article Title: Bromodomain and extra-terminal protein inhibitors modulate natural killer cell function and differentiation

doi: 10.1038/s41598-025-33437-1

Figure Lengend Snippet: BETi impact function of primary human NK cells. ( A ) Human primary NK cells isolated from healthy donors ( n = 4) unstimulated or stimulated with IL-12 + IL-15 for 24 h with prior (24 h) treatment with BETi (AZD5153 or CPI-203) compared to no drug treatment control. Bar graph of levels of IFNγ secreted into the cell culture supernatant measured by ELISA reported in pg/ml. ** P ≤ 0.01, * P ≤ 0.05, Ratio paired t-test. ( B ) Percentage of live CD14 − CD19 − cells and of total NK cells (live CD14 − CD19 − CD56 + CD16 + ) within the lymphocyte gate (Supplementary Fig. 2) in PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells for a further 5 h and analyzed. ( C ) Representative histogram overlays illustrating expression of granzyme B in NK cells in the indicated conditions, and bar graphs showing the median fluorescence intensity (MFI) of granzyme B staining in NK cells after culture of PBMCs from healthy donors ( n = 6) for 48 h in medium with or without IL-12 + IL-18 in the absence (open circles) or presence of BET inhibitors CPI-203 (black circles) or AZD5153 (blue circles), followed by co-culture for 5 h with CD4.221 cells in the presence of Brefeldin A. ( D – F ) Percentages of CD107a + ( D ), of IFN-γ + ( E ) and of TNF-α + cells ( F ) in total NK cells within PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells in the presence of Brefeldin A for a further 5 h and analyzed. In ( B – F ), each symbol represents data from a single donor; error bars indicate mean ± sem of n = 6 samples. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.00001; one way-ANOVA with Tukey’s post-test.

Article Snippet: Cells were stimulated with 10ng/ml recombinant IL-12 (130-129-720, Miltenyi) plus 100ng/ml recombinant IL-18 (B001-5, R&D Systems) and with 100nM CPI-203 (15479, Cayman Chemical) or 100nM AZD 5153 (15479, Cayman Chemical), or cultured in medium only for 48 h at 37 °C.

Techniques: Isolation, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Incubation, Expressing, Fluorescence, Staining, Co-Culture Assay

Journal: Scientific Reports

Article Title: Bromodomain and extra-terminal protein inhibitors modulate natural killer cell function and differentiation

doi: 10.1038/s41598-025-33437-1

Figure Lengend Snippet: BETi alter the transcriptome of human primary NK cells after activation. ( A ) Volcano plot of Log2 fold-change and significance of differentially expressed genes from BETi-treated (AZD5153 or CPI-203; 48 h treatment) compared to control treated following 24 h stimulation with IL-12/IL-15 determined by RNA-seq of primary human NK cells ( n = 4 per group). Blue dots represent significantly downregulated genes, and red dots represent significantly upregulated genes. Select individual genes are labeled. ( B ) Bar graphs of selected genes that encode PRF1, NKG2D, CD16, PD1, LAG3 and TIGIT showing normalized gene counts in each sample. Error bars indicate mean ± sem. Adjusted p value determined by DeSeq2 analysis with Bonferroni correction (*** P ≤ 0.001, * P ≤ 0.05). ( C ) Heatmap of Normalized Enrichment Scores (NES) calculated by Gene Set Enrichment Analysis (GSEA) for gene pathways that were negatively enriched with BETi treatment.

Article Snippet: Cells were stimulated with 10ng/ml recombinant IL-12 (130-129-720, Miltenyi) plus 100ng/ml recombinant IL-18 (B001-5, R&D Systems) and with 100nM CPI-203 (15479, Cayman Chemical) or 100nM AZD 5153 (15479, Cayman Chemical), or cultured in medium only for 48 h at 37 °C.

Techniques: Activation Assay, Control, RNA Sequencing, Labeling