Journal: Frontiers in immunology

Article Title: PCDHGA10 as a potential prognostic biomarker and correlated with immune infiltration in gastric cancer.

doi: 10.3389/fimmu.2024.1500478

Figure Lengend Snippet: FIGURE 4 Relationship between PCDHGA10 protein expression and tumor-infiltrating immune cells and immune checkpoints. (A-C) Multispectral composite of CD3, CD4, CD8, CD11b, CD66b, CD68, CK and DAPI, magnification ×200. (D) Four-color multispectral composite of LAG3, PD1, CK and DAPI, magnification ×200. Green: CK, blue: DAPI. (E) Correlation analysis of PCDHGA10 protein expression and tumor-infiltrating immune cells. (F) Correlation analysis of PCDHGA10 protein expression and immune checkpoints. DAPI: 4, 6-diamino-2-phenyl indole; CK, cytokeratin. * p < 0.05.

Article Snippet: This study used the following primary antibodies: rabbit antiPCDHGA10 (1:50, D1247, Biobyt), rabbit anti-CD11b antibody (1:100, 49420S, CST), rabbit anti-CD8 antibody (1:100, ab83278, Frontiers in Immunology 03 Abcam), rabbit anti-CD3 antibody (1:200, 85061S, CST), rabbit anti-CD4 antibody (1:200, ab133616, Abcam), mouse anti-Foxp-3 antibody (1:50, ab20034, Abcam), anti-LAG-3 antibody (1:50, ab52587, Abcam), anti-CD66b antibody (1:500, ab214175, Abcam), anti-cytokeratin antibody (1:8000, orb69073, Biobyt), anti-CD68 antibody (1:500, 797778S, CST).

Techniques: Expressing

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: Dopamine accumulates locally in HCC and promotes the proliferation, migration, and invasion of HCC cells in vitro . (A) DDC, MAOA, and dopamine levels in 30 matched non‐tumor and tumor tissues from HCC patients. (B) Analysis of the correlation between the levels of MAOA, DDC, and dopamine in 30 human HCC tissues. (C) Dopamine levels in HCC cell lines, hepatoblastoma Hep‐G2, and a normal liver cell line (Miha). (D) The growth rate of HCC cell lines, hepatoblastoma Hep‐G2, and a normal liver cell line (Miha) after treatment with dopamine (1 µM). (E) Migration and invasion in MHCC‐97H and SK‐Hep‐1. The data are presented as the mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: DDC, dopa decarboxylase. DRD1, dopamine receptor D1. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase. HCC, hepatocellular carcinoma. MAOA, monoamine oxidase A.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Migration, In Vitro

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: Expression of dopamine receptors in human HCC tissue. (A) Hierarchical clustering heat map showing the expression of mRNAs in HCC tissues and adjacent non‐tumor tissues by microarray analysis. The whole gene list and information are supplied in the Supplementary Table 5. (B) Waterfall plot of dopamine receptor D1 (DRD1) mRNA expression in 74 paired tumor and non‐tumor samples tested by qRT‐PCR. (C) DRD1 mRNA expression in tumor and adjacent non‐tumor tissues. (D) DRD1 expression in 7 paired tumor tissues and adjacent non‐tumor tissues tested by Western blot analysis and quantification of DRD1 protein levels. (E) DRD1 mRNA levels in HCC cell lines, hepatoblastoma Hep‐G2, and a normal liver cell line (Miha) were tested by qRT‐PCR. (F) DRD1 protein levels in HCC cell lines, hepatoblastoma Hep‐G2, and a normal liver cell line (Miha) were tested by Western blot analyses. * P < 0.05. Abbreviations: DRD1, dopamine receptor D1. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase. HCC, hepatocellular carcinoma.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Expressing, Microarray, Quantitative RT-PCR, Western Blot

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: DRD1 expression is an independent prognostic factor for HCC. (A) DRD1 immunohistochemical score in HCC tissues according to four staining intensity classes. (B) RFS curves of patients with HCC in relation to DRD1 protein expression by Kaplan‐Meier survival analysis. (C) OS curves of patients with HCC in relation to DRD1 protein expression by Kaplan‐Meier survival analysis. (D) Cox multivariate analysis of contributory factors to recurrence‐free survival and overall survival among 221 HCC patients after hepatectomy. Abbreviations: DRD1, dopamine receptor D1. HCC, hepatocellular carcinoma. OS, overall survival. RFS, relapse‐free survival.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: Correlation of DRD1 expression with the clinicopathological characteristics in 221 patients with HCC ¹

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Expressing

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: HCC can be regulated by interfering with DRD1 expression. (A) Western blot analysis confirmed the overexpression of DRD1 in Hep‐3B cells by stable transfection. (B) Western blot analysis confirmed the knockout of DRD1 expression in MHCC‐97H cells by stable transfection. (C) The upregulation of DRD1 expression significantly enhanced cell proliferation. (D) The downregulation of DRD1 expression weakened cell proliferation. (E) The upregulation of DRD1 expression significantly increased the colony numbers. (F) The downregulation of DRD1 expression decreased the colony numbers. (G) The upregulation of DRD1 expression significantly enhanced cell migration and invasion. (H) The downregulation of DRD1 expression weakened cell migration and invasion. The data are presented as the mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: DRD1, dopamine receptor D1. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase. HCC, hepatocellular carcinoma.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Expressing, Western Blot, Over Expression, Stable Transfection, Knock-Out, Migration

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: DRD1 activates the cAMP/PI3K/AKT/CREB pathway and Dopamine regulates HCC cell malignant behaviors via DRD1. (A) ELISA and Western blot analysis showed that the downregulation of DRD1 suppressed the cAMP/PI3K/AKT/CREB pathway, and the overexpression of DRD1 stimulated the cAMP/PI3K/AKT/CREB pathway. (B) Cell viability assay in sh‐DRD1 and sh‐NC cells with or without dopamine treatment (1 µM). (C) Colony formation assay in sh‐DRD1 and sh‐NC cells with or without dopamine treatment (1 µM). (D) Migration and invasion in sh‐DRD1 and sh‐NC cells with or without dopamine treatment (1 µM). (E) The cAMP/PI3K/AKT/CREB pathway was affected by treatment with dopamine (1 µM). The data are presented as the mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: DRD1, dopamine receptor D1. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase. HCC, hepatocellular carcinoma. cAMP, 3',5'‐cyclic adenosine monophosphate. PI3K, phosphoinositide 3‐kinase. AKT, protein kinase B. CREB, cAMP response element‐binding protein.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Over Expression, Viability Assay, Colony Assay, Migration, Binding Assay

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: SCH23390 plays a tumor‐suppressive role both in vitro and in vivo . (A) Cell viability assay in HCC cell lines, hepatoblastoma Hep‐G2, and Miha cells after treatment with SCH23390 (100 µM, a selective DRD1 antagonist). (B) Migration and invasion of MHCC‐97H and SK‐Hep‐1 cells after treatment with SCH23390. (C) SCH23390 (0.1 mg/kg) restrained tumor growth (compared with the control) in the xenograft tumor model. The data are presented as the mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: DRD1, dopamine receptor D1. HCC, hepatocellular carcinoma.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques: In Vitro, In Vivo, Viability Assay, Migration, Control

Journal: Cancer Communications

Article Title: Increased dopamine and its receptor dopamine receptor D1 promote tumor growth in human hepatocellular carcinoma

doi: 10.1002/cac2.12103

Figure Lengend Snippet: Model depicting the possible mechanisms of the dopaminergic system locally contributed to HCC progression via DRD1.

Article Snippet: The DRD1 antibody (NBP2‐16213) was purchased from Novus Biological (CO, USA).

Techniques:

Journal:

Article Title: Essential Role of AKT-1/Protein Kinase B? in PTEN-Controlled Tumorigenesis

doi: 10.1128/MCB.22.11.3842-3851.2002

Figure Lengend Snippet: Growth potential, cell cycle profile, and analysis of cell cycle regulators in Pten/Akt clones. (A) Growth curve of indicated ES clones. (B) Growth competition assay. Pten−/−;Akt−/− ES cells were cocultivated with equal numbers of WT or Pten−/− cells. DNA was extracted from the cocultures after each passage and was analyzed with Southern analysis to determine the ratio of WT Akt allele (23 kb) to mutant Akt allele (15 kb). Lanes 1 and 2, WT and Pten−/−;Akt−/− cells cultured alone. Lanes 3 to 6, Pten−/−;Akt−/− ES cells cocultured with WT cells at different passages (p1 to p4). Lanes 7 to 10, Pten−/−;Akt−/− ES cells cocultured with Pten−/− cells at different passages (p1 to p4). KO, knockout. (C) Western blot analysis of cell cycle regulators. Total cell lysates (20 μg) were separated on sodium dodecyl sulfate-polyacrylamide gels and blotted with cell cycle inhibitors p27KIP1 and p16INK1 as well as cyclins A, D1, and E. Vinculin (vinc) was used on each blot as a loading control. (D) Cell cycle analysis. ES cells were synchronized at M phase with Colcemid treatment followed by replating in Colcemid-free medium to allow reentry into the cell cycle. FACS analysis was performed on each clone at different time points (2 and 3 h), and the percentages of cells remaining in M phase are shown here.

Article Snippet: Antibodies specific for ERK, p27 KIP1 (sc-528), cyclin D1 (R-124), p21 CIP1/WAF1 (sc-397), cyclin A, and mouse cyclin E (sc-481) were obtained from Santa Cruz Biotechnology.

Techniques: Clone Assay, Competitive Binding Assay, Mutagenesis, Cell Culture, Knock-Out, Western Blot, Cell Cycle Assay

Journal: bioRxiv

Article Title: Microglia-mediated synaptic pruning in the nucleus accumbens during adolescence: A preliminary study of the proteomic consequences and putative female-specific pruning target

doi: 10.1101/2023.05.02.539121

Figure Lengend Snippet: In remaining tissue (unpooled) that underwent proteomic analysis ( , ) we conducted ELISAs to validate potential female-specific pruning targets. (A) Inhibiting pruning in the NAc significantly decreased Lynx1 expression in females, but not males. n =7–8/sex/condition. (B) The Lynx1 target, NAChR, was not regulated by NIF treatment in either sex. n =7–8/sex/condition. (C) In remaining, but underpowered sample sizes ( n =3–5/sex/condition), we observed the expected increase in D1r levels after inhibiting NAc pruning in males, but not females. This is consistent with our previously published results. In each histogram, horizontal lines are average and vertical lines are standard error of the mean. * p <0.05.

Article Snippet: Lynx1 ELISA was purchased from mybiosource.com (#MBS9902329), NAChR ELISA was purchased from LSBio (#LS-F37436), and D1r ELISA was purchased from Novus Biologicals (NBP2–67935).

Techniques: Expressing

Journal: Translational Cancer Research

Article Title: DRD1 and DRD4 are differentially expressed in breast tumors and breast cancer stem cells: pharmacological implications

doi: 10.21037/tcr-22-783

Figure Lengend Snippet: Expression of DRs breast cancer cell lines. (A) mRNA expression of DRDs in 62 breast cancer cell lines form the Cancer Cell Line Encyclopedia. MCF-7 and MDA-MB-231 cell lines are shown in magenta and red, respectively. (B,C) Protein quantification of DRD1, DRD2, and DRD4 in MCF-7 (B) and MDA-MB-231 (C) by flow cytometry. Graphs in (B) and (C) show MFI (average ± SEM) from 2–3 independent experiments. **, P<0.05 (Student’s t- test). DR, dopamine receptor; MFI, mean fluorescence intensity; SEM, standard error of the mean; TPM, transcript per million.

Article Snippet: Expression of DRs was analyzed using anti-human DRD1 Alexa Fluor 405 (FAB8276V, R&D Systems), anti-human DRD2 Alexa Fluor 647 (sc-5303, Santa Cruz Biotechnology), or anti-human DRD4 PE (sc-136169, Santa Cruz Biotechnology).

Techniques: Expressing, Flow Cytometry, Fluorescence