Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: RNA sequencing data analysis in nax ( n = 5) and wild–type (wt) ( n = 6) mice. Data analysis shows dopamine receptor D1 ( Drd1, A ), Drd3 ( C ), and Drd4 ( D ) are significantly increased in the nax cerebellum. Although an increase in Drd2 ( B ) and Drd5 ( E ) is apparent, statistical significance was not reached. The data in the bar graph are presented as the mean ± SEM, and statistical analysis was performed using an unpaired t –test (* p < 0.05 and ** p < 0.01). RPKM: Reads Per Kilobase of transcript per Million mapped reads.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: RNA Sequencing Assay

Journal: International Journal of Molecular Sciences

Article Title: Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia ( NAX )) Mouse

doi: 10.3390/ijms21082914

Figure Lengend Snippet: Frontal sections of wt and nax mouse cerebella: DRD1 expression at P5 and P17. ( A ) Immunoperoxidase staining for DRD1 in the wt cerebellum on P5 shows the immunoreactivity in the Purkinje cell layer (Pcl) and white matter (wm) but not in the external germinal zone (egz) and granular layer (gl). ( B ) Immunoperoxidase staining for DRD1 in the nax cerebellum shows similar immunoreactivity as in the wt littermates at P5. ( C ) DRD1 immunostaining of the frontal sections of the wt cerebellum at P17 shows immunoreactivity in the Purkinje cell (PC) somata, the Pcl, and dendrites in the molecular layer (ml) and in the gl but not in the wm. ( D ) DRD1 immunostaining of the frontal sections of the nax cerebellum on P17 shows immunoreactivity in the PCs in the ml/Pcl. ( E ) Western blot analysis of whole cerebellar lysates revealed no significant differences in DRD1 expression between wt and nax cerebella at P5 and P17 (wt: n = 3 and nax : n = 3). The data in the bar graphs are presented as the means of three independent experiments ± SEM, and statistical analysis was performed using a two–way ANOVA. P; postnatal. Scale bars: 100 μm in A, B, C, and D.

Article Snippet: D1: rabbit polyclonal anti–D1 dopamine receptor (TA328798, anti–Drd1, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA); D2: rabbit polyclonal anti–D2 dopamine receptor (TA328800, anti–Drd2, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 2 (DR2); D3: rabbit polyclonal anti–D3 dopamine receptor (TA328800, anti–Drd3, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 3 (DR3); D4: rabbit polyclonal anti–D4 dopamine receptor (TA321202, anti–DRD4, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor D4 (DRD4); and D5: rabbit polyclonal anti–D5 dopamine receptor (TA328802, anti–Drd5, diluted 1:1000; OriGene Biotech Co., Rockville, MD, USA), produced against recombinant rat dopamine receptor 5 (DR5).

Techniques: Expressing, Immunoperoxidase Staining, Immunostaining, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Effects of a Chlorogenic Acid-Containing Herbal Medicine (LAS NB ) on Colon Cancer

doi: 10.1155/2021/9923467

Figure Lengend Snippet: Control and LAS NB xenograft models. (a) Hematoxylin-eosin staining assay results are shown. Compared with the control group, the LAS NB group showed increased colon cell apoptosis. (b) CD34, EGF, and VEGF expression and TUNEL-positive cells were assessed by immunohistochemistry in the tumor microenvironment. Magnification, 400x ( p < 0.05). (c) Western blot results are shown. The expression of RTK kinase (EGF and VEGF), NF- κ B proteins (NF- κ B p65, NF- κ B p50, and TGF- β ), and MAPK (p-ERK and t-ERK) in the xenograft model (C1), CT26 cells (C2), and HCT15 cells (C3). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.005.

Article Snippet: Subsequently, the following steps were performed: blocking of endogenous peroxidase activity in 3% H 2 O 2 solution, unmasking of the antigenic epitope with citrate buffer, addition of blocking buffer for blocking, and addition of primary antibody VEGF (Bioss, BS1665R, 1 : 1000), CD34 (Bioss, BS5085R, 1 : 1000), and EGF (Bioss, BS1007R, 1 : 1000) and a secondary antibody (horseradish peroxidase-labeled goat rabbit IgG [H + L]; Beyotime, A0208).

Techniques: Staining, Expressing, TUNEL Assay, Immunohistochemistry, 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: 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: Nature Communications

Article Title: Cell-type specific profiling of histone post-translational modifications in the adult mouse striatum

doi: 10.1038/s41467-022-35384-1

Figure Lengend Snippet: A Immunohistochemistry displaying DAPI/GFP/A2a (left) and D1 (right) from the striatum of SUN1-GFP;A2a-CRE + animals. n = 3/group. Arrows indicate co-localization of all three markers. B Immunohistochemistry displaying DAPI/GFP/Drd1 (left) and A2a (right) from the striatum of SUN1-GFP; D1-CRE + animals. n = 3/group. Arrows indicate co-localization of all three markers. These experiments were replicated 3 times with similar results. C INTACT schematic created at Biorender.com. D Total (%) of Cre + cells in striatum. Image depicts total nuclei isolation ( n = 2/cell type and antibody) E Yield (%) and specificity (%) of GFP + nuclei. Image depicts GFP + nuclei isolation. n = 2/cell type and antibody. F mRNA validation for INTACT Sun1-GFP; A2a-Cre + vs bulk (Cre- nuclei); n = 5, unpaired two-tailed t test, A2a Nuclei, Drd1: (t (10) = 9.871, p < 0.001); A2a: (t (10) = 4.129, p = 0.006.), normalized to GAPDH. Data are presented as mean values + /- SEM * P < 0.05. G mRNA validation for INTACT Sun1-GFP; D1-Cre + vs bulk (Cre- nuclei); n = 5, unpaired two-tailed t test, D1 Nuclei, D1: (t (10) = 2.920, p = 0.0278); A2a: (t (10) = 3.421, p = 0.012), normalized to GAPDH. Data are presented as mean values + /- SEM. * P < 0.05. Source data are provided as a Source Data file.

Article Snippet: Sections were blocked for 90 minutes at room temperature with 10% Normal Donkey Serum and 0.2% Triton X-100 in PBS and then incubated with 0.2% Triton X-100 in PBS with the following antibodies overnight at 4 °C: rabbit anti-Drd1 (1:500, Bioss USA BSM-52920R) or rabbit anti-A2a (1:200, Fisher Scientific PA1-042), co-incubated with goat anti-GFP (1:500, Rockland, 600-101-215).

Techniques: Immunohistochemistry, Isolation, Two Tailed Test

Journal: Nature Communications

Article Title: Cell-type specific profiling of histone post-translational modifications in the adult mouse striatum

doi: 10.1038/s41467-022-35384-1

Figure Lengend Snippet: A-C ) A2a and D1 specific H3K4me3 and H3K27me3 modifications around known A2a-specific cell-type markers ( A ) A2a, (B) Drd2, and ( C ) Penk . D-F) A2a and D1 specific H3K4me3 and H3K27me3 modifications around known D1-specific cell-type markers ( D ) Drd1 , ( E ) Pdyn, and ( F ) Tac1 . ( G ) Peaks at A2a and ( H ) D1 upregulated genes; cell type-specific genes defined by |log2fold change | > 1 and adj P < 0.05. ( I ) A2a H3K4me3 signal, ( J ) A2a H3K27me3 signal, ( K ) D1 H3K4me3 signal, ( L ) and D1 H3K27me3 signal centered on transcription start site (TSS). For each cell-type and hPTM, genes are categorized into 3 groups: high (FPKM > = 10), medium (1 < = FPKM < 10), and low (FPKM < 1) based on published MSN-specific Ribo-Tag . Signal is normalized to total mapped reads: Count Per Million (CPM). M ) A2a and D1 H3K4me3 signal centered on the TSS of A2a-specific genes, ( N ) A2a and D1 H3K4me3 signal centered on the TSS of D1-specific genes, ( O ) A2a and D1 H3K27me3 signal centered on the TSS of A2a-specific genes, ( P ) A2a and D1 H3K27me3 signal centered on the TSS of D1-specific genes based on published MSN-specific Ribo-Tag . Signal is normalized by variance across transcriptionally constant genes (quantile normalization). A2a and D1-specific genes are defined as q -value <0.05 and | log2 FC | > 1.

Article Snippet: Sections were blocked for 90 minutes at room temperature with 10% Normal Donkey Serum and 0.2% Triton X-100 in PBS and then incubated with 0.2% Triton X-100 in PBS with the following antibodies overnight at 4 °C: rabbit anti-Drd1 (1:500, Bioss USA BSM-52920R) or rabbit anti-A2a (1:200, Fisher Scientific PA1-042), co-incubated with goat anti-GFP (1:500, Rockland, 600-101-215).

Techniques:

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