Journal: Journal for Immunotherapy of Cancer

Article Title: Pharmacologic targeting of the dopamine D2 receptor impacts the efficacy of immune checkpoint blockade in melanoma

doi: 10.1136/jitc-2025-014080

Figure Lengend Snippet: Graphical abstract. Figure created in BioRender. BRC, bromocriptine; D2R, D2 receptor; HAL, haloperidol; ICI, immune checkpoint inhibitor; IL, interleukin; MCP, metoclopramide; MHC, major histocompatibility complex; PRL, prolactin.

Article Snippet: Samples were loaded in 10% precast polyacrylamide gels (Bio-Rad, cat. #4561036), run at 100 V in Tris/Glycine/sodium dodecyl sulfate (SDS) buffer (Bio-Rad 1610732), transferred to polyvinylidene fluoride (PVDF) membrane (Bio-Rad, cat. #1704272), blocked for 1 hour with 5% dry milk in tris buffered saline-Tween (TBS-T), and probed for PRL (R&D, cat. #AF1445), D2R (Proteintech, cat. #55084-1-AP), and β-Actin-HRP (BioLegend, cat. #664803) in blocking buffer.

Techniques: Immunopeptidomics

Journal: Journal for Immunotherapy of Cancer

Article Title: Pharmacologic targeting of the dopamine D2 receptor impacts the efficacy of immune checkpoint blockade in melanoma

doi: 10.1136/jitc-2025-014080

Figure Lengend Snippet: Tumor immune microenvironment in prolactin-locus ICI non-responder and responder models. ( A ) Experimental scheme. B16F0-bearing (CC51xB6)F1 and B6 mice received 100 µg αCTLA-4 and 200 µg αPD-1 on days 3, 6, and 10 after tumor inoculation. On day 13 after tumor inoculation, tumors were enriched for CD45+ TILs then processed for scRNA-seq. ( B ) UMAP and ( C ) composition analysis showing the breakdown of B6U, B6T, CC51U, CC51T annotated cell types overlaying the UMAP ( B ) and proportions of each cell type cluster. For each experimental condition, cell proportions (Y axis) were calculated by dividing the number of cells of a given type by the total number of cells. B6U, B6 untreated; B6T, B6 ICI-treated; CC51U, (CC51xB6)F1 untreated; CC51T, (CC51xB6)F1 ICI-treated. Cell types annotated manually using canonical markers. ( D ) Expression levels of M1 and M2 markers in the MΦ cluster. ( E ) Volcano plot highlighting DEGs in the CD20− CD8+ T cluster between B6T and CC51T. Significant DEG (padj≤0.05, |log2FC | ≥ 1) were determined using FindMarkers and are colored (red=upregulated in B6T; purple=upregulated in CC51T). ( F ) Expression levels of Prlr, Drd2, Htr3a and Htr4 in all clusters. αCTLA-4, anti-cytotoxic T-lymphocyte-associated protein 4; αPD-1, anti-programmed cell death protein-1; B, B cells; B16, B16 F0 cells; CD4+ T, CD4+ T cells; CD20- CD8+ T, CD8+ T cells; CD20+ CD8+ T, CD20+ CD8 T cells; cDC1, conventional dendritic cells, type I; DEGs, differentially expressed genes; Drd2, dopamine D2 receptor; Htr3a, serotonin 5HT3 receptor; Htr4, 5HT4 receptors; ICI, immune checkpoint inhibitor; MΦ, macrophages; MDSC, myeloid-derived suppressor cells; Mono, monocytes; NK T, natural killer T cells; Prlr, prolactin receptor; scRNA-seq, single-cell RNA sequencing; TILs, tumor-infiltrating leukocytes.

Article Snippet: Samples were loaded in 10% precast polyacrylamide gels (Bio-Rad, cat. #4561036), run at 100 V in Tris/Glycine/sodium dodecyl sulfate (SDS) buffer (Bio-Rad 1610732), transferred to polyvinylidene fluoride (PVDF) membrane (Bio-Rad, cat. #1704272), blocked for 1 hour with 5% dry milk in tris buffered saline-Tween (TBS-T), and probed for PRL (R&D, cat. #AF1445), D2R (Proteintech, cat. #55084-1-AP), and β-Actin-HRP (BioLegend, cat. #664803) in blocking buffer.

Techniques: Expressing, Derivative Assay, Single Cell, RNA Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Grapefruit By-Products as a Sustainable Source of Bioaccessible Polyphenols with In Vitro Neuroprotective Potential

doi: 10.3390/ijms27073140

Figure Lengend Snippet: Digested GDSE solution stimulates the synthesis of genes related to neuroprotection in SH-SY5Y cells. mRNA expression levels of BDNF and DRD2 after incubation of SH-SY5Y cells in the presence of a non-cytotoxic dilution (1/16, corresponding to a concentration of 0.625 mg mL −1 ) of the indicated sample digestions during a 4-h period ( n ≥ 6). Data are presented as mean ± SEM of at least three independent experiments. Two-tailed Student’s t -test adjusted for multiple comparisons using the Holm-Šídák method. Significance is indicated as: ** p ≤ 0.0201.

Article Snippet: Specific biomarkers were selected for each cell model and amplified using TaqMan Gene Expression Assays (Thermo Fisher Scientific, Waltham, MA, USA), including GAPDH (glyceraldehyde-3-phosphate dehydrogenase; Hs02758991_g1), BDNF (Hs02718934_s1), DRD2 (Hs00241436_m1), BCL2 (Hs00608023_m1), DLG4 (Hs01555373_m1), and SYN1 (Hs00199577_m1).

Techniques: Expressing, Incubation, Concentration Assay, Two Tailed Test

Journal: bioRxiv

Article Title: D2 autoreceptors gate vulnerability to cocaine use disorder

doi: 10.64898/2026.03.10.710882

Figure Lengend Snippet: a , Schematic of the principal contributors to striatal D2/3 ligand binding: postsynaptic D2 heteroreceptors on medium spiny neurons (MSNs; red) and presynaptic D2 autoreceptors on dopamine axons (blue). b , Breeding strategy generating littermates with cell-type–specific Drd2 haploinsufficiency. Genotypes: control (Drd2 loxP/wt ; black), autoD2KD (Drd2 loxP/wt ; Dat IRES-Cre/wt ; blue), MSN-D2KD (Drd2 loxP/wt ; Adora2a-Cre +/– ; red), and double-D2KD (Drd2 flox/wt ; Adora2a-Cre +/– ; Dat IRES-Cre/wt ; purple). Bottom, RT–qPCR quantification of Drd2 mRNA (ΔΔCt) in frontal cortex (open), nucleus accumbens (NAc; dark), and dorsal striatum (light) (n = 5 control; n = 3–5 autoD2KD; n = 5 MSN-D2KD; n = 3–5 double-D2KD). c, e , Representative autoradiographs of [3H]raclopride binding (c) and [3H]SCH23390 binding (e), including nonspecific binding. Color scale indicates nCi/g. Autoradiography was performed in two independent cohorts, each with its own control group; representative control images are therefore shown twice. d, f , Left, schematic of striatal regions of interest used for quantification. Right, specific binding in dorsal striatum and NAc, normalized to the mean of the corresponding control region, for [3H]raclopride (d) and [3H]SCH23390 (f). Symbols denote individual mice; bars show mean ± s.e.m. For binding quantification, n = 3–9 mice per genotype; 8–12 sections per mouse; 24–30 ROIs per animal. *P ≤ 0.05; #0.1 ≥ P > 0.05.

Article Snippet: Relative dopamine D2 receptor (Oa04895884_m1 TaqMan, Catalog #: 4351372 ) and beta-actin probe (Mm01205647_g1 TaqMan, Catalog #: 4331182) mRNA expression were determined with TaqMan Gene Expression Assays (Life Technologies) using a StepOnePlus Real-Time PCR system (Applied Biosystems).

Techniques: Ligand Binding Assay, Control, Quantitative RT-PCR, Binding Assay, Autoradiography