Journal: Frontiers in Pharmacology

Article Title: LEAP2 Impairs the Capability of the Growth Hormone Secretagogue Receptor to Regulate the Dopamine 2 Receptor Signaling

doi: 10.3389/fphar.2021.712437

Figure Lengend Snippet: LEAP2 impairs the basal reduction of Ca V 2.2 currents by GHSR and D2R coexpression and LEAP2 ameliorates the ability of GHSR to impair dopamine-induced inhibition of Ca V 2.2 currents. (A) Representative traces (left) of Ca V 2.2 current (I CaV2.2 ) from HEK293T cells cotransfected with Ca V 2.2, Ca V β 3 , Ca V α 2 δ 1 and either D2R (+D2R, n = 18), GHSR (+GHSR, n = 21) or GHSR and D2R (+D2R +GHSR, n = 17) pre-incubated or not with 1 µM SPA (+D2R +GHSR +SPA, n = 10) or 0.1 µM LEAP2 (+D2R +GHSR +LEAP2, n = 21) in a 0.1 GPCR:Ca V 2.2 molar ratio. Bars (right) represent averaged I CaV2.2 levels for each condition. Statistical significance was evaluated by Kruskal-Wallis and Dunn’s post-test. (B) Representative traces and time courses (left) of Ca V 2.2 current (I CaV2.2 ) from HEK293T cells cotransfected with Ca V 2.2, Ca V β 3 , Ca V α 2 δ 1 and D2R (+D2R, n = 9) or GHSR and D2R pre-incubated or not (+D2R +GHSR, n = 7) with 1 µM SPA (+D2R +GHSR +SPA, n = 5) or 0.1 µM LEAP2 (+D2R +GHSR +LEAP2, n = 8) in control condition and after dopamine application (10 µM, +DA); 0.1 GPCR:Ca V 2.2 molar ratio. Bars (right) represent averaged I CaV2.2 inhibition by 10 µM dopamine application for each condition. Statistical significance was evaluated by Kruskal-Wallis and Dunn’s post-test (versus +D2R). The test-pulse protocol consisted in square pulses applied from −100 to +10 mV for 30 ms every 10 s.

Article Snippet: HEK293T cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) with 10% fetal bovine serum (Internegocios) and transfected with plasmids containing Ca V 2.2 (#AF055477), auxiliary subunits Ca V β 3 (#M88751) and Ca V α 2 δ 1 (#AF286488), GFP-containing plasmid (to identify transfected cells) with or without GHSR-containing plasmid (#AY429112) and/or D2R-containing plasmid (MG226860-Origene).

Techniques: Inhibition, Incubation

Journal: Frontiers in Pharmacology

Article Title: LEAP2 Impairs the Capability of the Growth Hormone Secretagogue Receptor to Regulate the Dopamine 2 Receptor Signaling

doi: 10.3389/fphar.2021.712437

Figure Lengend Snippet: LEAP2 (1-14) impairs GHSR modulation of D2R signaling. (A) Representative traces (left) of Ca V 2.2 current (I CaV2.2 ) from HEK293T cells cotransfected with Ca V 2.2, Ca V β 3 , Ca V α 2 δ 1 and either D2R (+D2R, n = 14) or GHSR and D2R (+D2R +GHSR, n = 16) pre-incubated or not with 0.1 µM LEAP2 (1–14) [+D2R +GHSR +LEAP2 (1–14), n = 26] or 0.1 µM LEAP2 (15–40) [+D2R +GHSR +LEAP2 (15–40), n = 19] in a 0.1 GPCR:Ca V 2.2 molar ratio. Bars (rigth) represent averaged I CaV2.2 levels for each condition. Statistical significance was evaluated by Kruskal-Wallis and Dunn’s post-test (versus +D2R condition). (B) Representative traces and time courses (left) of Ca V 2.2 current (I CaV2.2 ) from HEK293T cells cotransfected with Ca V 2.2, Ca V β 3 , Ca V α 2 δ 1 and D2R (+D2R, n = 8) or GHSR and D2R pre-incubated or not (+D2R+GHSR, n = 6) with 0.1 µM LEAP2 (1–14) [+D2R +GHSR +LEAP2 (1–14), n = 10] or 0.1 µM LEAP2 (15–40) [+D2R +GHSR +LEAP2 (15–40), n = 5] in control condition and after dopamine application (10 µM, +DA); 0.1 GPCR:Ca V 2.2 molar ratio. Bars (right) represent averaged I CaV2.2 inhibition by 10 µM dopamine application for each condition. Statistical significance was evaluated by Kruskal-Wallis and Dunn’s post-test (versus +D2R). The test-pulse protocol consisted in square pulses applied from −100 to +10 mV for 30 ms every 10 s.

Article Snippet: HEK293T cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) with 10% fetal bovine serum (Internegocios) and transfected with plasmids containing Ca V 2.2 (#AF055477), auxiliary subunits Ca V β 3 (#M88751) and Ca V α 2 δ 1 (#AF286488), GFP-containing plasmid (to identify transfected cells) with or without GHSR-containing plasmid (#AY429112) and/or D2R-containing plasmid (MG226860-Origene).

Techniques: Incubation, Inhibition

Journal: Frontiers in Pharmacology

Article Title: LEAP2 Impairs the Capability of the Growth Hormone Secretagogue Receptor to Regulate the Dopamine 2 Receptor Signaling

doi: 10.3389/fphar.2021.712437

Figure Lengend Snippet: Impact of LEAP2 on GHSR structure and dopamine-mediated Gi activation. (A) XL255 emission intensity after Tb-cryptate excitation of proteoliposomes containing Tb-cryptate labeled GHSR and XL255-labeled D2R in absence of ligand (No ligand) or in presence of 10 µM SPA (+SPA) or LEAP2 (1–12) [+LEAP2 (1–12)]. Statistical significance was evaluated by One Way ANOVA and Tukey’s post-test. (B) Proximity ratio changes induced by 10 µM of SPA (+SPA) or LEAP2 (1–12) [+LEAP2 (1–12)] calculated from the FRET signal between the fluorophores in TM1 and TM6 of GHSR assembled into lipid nanodiscs either as a homomer (+GHSR) or a heteromer (+GHSR +D2R). (C) AF-488 emission intensity after AF-350 excitation. Gαi and Gαq were labeled at their N terminus with AF-350 and AF-488, respectively, and fluorescence was measured in the presence of the labeled G proteins, the GHSR-D2R heteromer in lipid nanodiscs and 10 µM ghrelin (+Ghrelin), 10 µM dopamine (+DA), or 10 µM dopamine in the absence or in the presence of either 10 µM SPA (+DA +SPA) or LEAP2 (1–12) [+DA +LEAP2 (1–12)]. (D) GTPγS binding to Gα i1 in Gα i1 β 1 γ 2 catalyzed by the GHSR-D2R heteromer in the presence of 10 µM dopamine (DA) and in absence or in the presence of either 10 µM SPA or LEAP2 (1–12). GTPγS binding to Gα i1 catalyzed under the same conditions by the D2R homomer in the presence of 10 µM dopamine (DA) is given for comparison. The species considered are schematically depicted in all cases (red: GHSR, blue:D2R, green: G protein). Data in ( A – C ) is mean ± SD of three experiments.

Article Snippet: HEK293T cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) with 10% fetal bovine serum (Internegocios) and transfected with plasmids containing Ca V 2.2 (#AF055477), auxiliary subunits Ca V β 3 (#M88751) and Ca V α 2 δ 1 (#AF286488), GFP-containing plasmid (to identify transfected cells) with or without GHSR-containing plasmid (#AY429112) and/or D2R-containing plasmid (MG226860-Origene).

Techniques: Activation Assay, Labeling, Fluorescence, Binding Assay

Journal: Endocrine Pathology

Article Title: Clinical and Molecular Characteristics of Gonadotroph Pituitary Tumors According to the WHO Classification

doi: 10.1007/s12022-023-09794-w

Figure Lengend Snippet: Gene expression data obtained in 54 GnPT: correlation matrix

Article Snippet: Two commercial antibodies were tested for D2R expression: a monoclonal antibody (mab9266–100, R&D system, distributed by Aurogene, Italy, referred to as D2R-mAb, dilution 1:100) and a polyclonal antibody (AB5084P, Sigma-Aldrich, distributed by Sial, Italy, referred to as D2R-pAb, dilution 1:500), the latter being previously used in this indication [ , ].

Techniques: Expressing

Journal: Endocrine Pathology

Article Title: Clinical and Molecular Characteristics of Gonadotroph Pituitary Tumors According to the WHO Classification

doi: 10.1007/s12022-023-09794-w

Figure Lengend Snippet: Bio-clinical and molecular characteristics of GnPT according to the presence or the absence of gonadotropin immunostaining

Article Snippet: Two commercial antibodies were tested for D2R expression: a monoclonal antibody (mab9266–100, R&D system, distributed by Aurogene, Italy, referred to as D2R-mAb, dilution 1:100) and a polyclonal antibody (AB5084P, Sigma-Aldrich, distributed by Sial, Italy, referred to as D2R-pAb, dilution 1:500), the latter being previously used in this indication [ , ].

Techniques: Expressing, Immunohistochemistry

Journal: Endocrine Pathology

Article Title: Clinical and Molecular Characteristics of Gonadotroph Pituitary Tumors According to the WHO Classification

doi: 10.1007/s12022-023-09794-w

Figure Lengend Snippet: D2R gene expression in GnPT and functional lactotroph tumors. No significant difference in D2R expression was found between functional lactotroph tumors (PRL) and GnPT, or between FSH/LH and pSF1 phenotypes ( A ). In contrast, D2R transcripts were significantly lower in recurrent GnPT as compared to non-recurrent cases ( B )

Article Snippet: Two commercial antibodies were tested for D2R expression: a monoclonal antibody (mab9266–100, R&D system, distributed by Aurogene, Italy, referred to as D2R-mAb, dilution 1:100) and a polyclonal antibody (AB5084P, Sigma-Aldrich, distributed by Sial, Italy, referred to as D2R-pAb, dilution 1:500), the latter being previously used in this indication [ , ].

Techniques: Expressing, Functional Assay

Journal: Endocrine Pathology

Article Title: Clinical and Molecular Characteristics of Gonadotroph Pituitary Tumors According to the WHO Classification

doi: 10.1007/s12022-023-09794-w

Figure Lengend Snippet: D2R immunostaining in controls and representative examples of GnPT using a monoclonal (D2R-mAb) or a polyclonal (D2R-pAb) antibody. In the brain tissue ( A , B ) (×200 magnification; inset at ×400), D2R staining was essentially cytoplasmic. In normal pituitary gland fragments, nuclear ( C ) or cytoplasmic with occasional membrane ( D ) D2R staining was observed (×200 magnification; inset at ×400). Similar results were obtained on functional lactotroph tumors ( E , F ) (×200 magnification; inset at ×400). Representative examples of D2R immunostaining in GnPT are also shown ( G , H ) using the mAb ( G ) nuclear staining which was observed in a normal juxta-tumoral pituitary fragment (indicated by a star) and in scattered neoplastic cells (inset) (×100 magnification; inset at ×400), whereas cytoplasmic staining was focally observed with the pAb (H) (×400 magnification)

Article Snippet: Two commercial antibodies were tested for D2R expression: a monoclonal antibody (mab9266–100, R&D system, distributed by Aurogene, Italy, referred to as D2R-mAb, dilution 1:100) and a polyclonal antibody (AB5084P, Sigma-Aldrich, distributed by Sial, Italy, referred to as D2R-pAb, dilution 1:500), the latter being previously used in this indication [ , ].

Techniques: Immunostaining, Staining, Membrane, Functional Assay

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 is transcriptionally downregulated through promoter methylation in BrCa. ( A ) Heatmap of top 30 differentially expressed genes based on RNA-seq. Tissues derived from nor mal breast tissues and BrCa tissues were applied for RNA-seq analysis. Log2FC transformed and normalized values were used. ( B ) IHC staining of DRD2 in normal breast tissues and BrCa tissues. Bars, 60 μm. ( C ) Expression and methylation of DRD2 based on TCGA database. Data are presented as mean ± SD; p -value was calculated using two-tailed Student's t test. p < 0.0001. ( D ) Online database Kmplot was used to analyze the effects of DRD2 on the overall prognosis (left) of BrCa patients, the survival times of BrCa patients featured HER2-positive (middle), and the survival times of BrCa patients featured Luminal A (right). ( E ) mRNA expression (RT-PCR) and promoter methylation (MSP) analysis of DRD2 in BrCa cells lines and mammary epithelial cell lines were all detected. ( F ) mRNA expression of DRD2 after Aza treatment was determined by qRT-PCR. MDA-MB231 and BT549 were treated with Aza for 3 d. BrCa cells without Aza treatment were used as controls. Data are presented as mean ± SD; p -value was calculated using two-tailed Student's t test. ***, p < 0.001. BN, normal breast; BrCa: Breast cancer; Ctrl, Control; MSP, methylation-specific PCR; M, methylated; U, unmethylated.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Methylation, RNA Sequencing Assay, Derivative Assay, Transformation Assay, Immunohistochemistry, Expressing, Two Tailed Test, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: Relationship between clinicopathological features and DRD2 promoter methylation in breast cancer (TCGA)

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Methylation

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 expression inhibits BrCa cells tumorigenesis in vitro and in vivo . ( A and B ) Confirming ectopic DRD2 mRNA expression by RT-PCR (A) and protein expression by WB (B). ( C ) Measurement of proliferation in Vector- and DRD2-transfected BrCa cells by CCK8 assay. ( D and E ) Histogram statistics of colony formation (D) and soft agar formation assay (E) to determine proliferation rates. ( F ) Histogram statistics showing analysis of apoptosis determined by AO/EB assay. ( G ) Histogram statistics of cell cycle distribution by FC. ( H ) Histogram statistics of proliferation rates in BrCa cells. CCK8 was performed to analyze effect of 891 DRD2 expression on chemosensitivity of BrCa cells to PTX. DMSO was used as controls. ( I and J ) Histogram showing effects of DRD2 on metastatic abilities in wound-healing (I) and Transwell® assay (J). Transwell® coated without (left) or with (right) Matrigel were applied to detecting migrative or invasive abilities of BrCa cells. ( K ) The volume and weight measurements of subcutaneous tumor model in BALB/c mice (8 mice per group). Volume = length × width2 × 0.5. DRD2/4T1 cells were used, and vector-transfected 4T1 cells were used as controls. Data are presented as mean ± SD; P-value was calculated using two-tailed Student's t test. *, p < 0.05; **, p < 0.01; ***, p < 0.001. PTX, Paclitaxel. AO/EB, acridine orange/ethidium bromide.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Expressing, In Vitro, In Vivo, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Transfection, CCK-8 Assay, Tube Formation Assay, AO/EB Assay, Transwell Assay, Two Tailed Test

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 reprograms Mφ towards M1 phenotype and downregulates IL-6 and IL-10. ( A ) IHC staining on serial sections of tissues from patients with BrCa. M1 markers, iNOS and CD68; M2 markers, CD206 and CD163. Bars, 80 μm. ( B and C ) qRT-PCR was used to detect M1 and M2 Mφ markers after co-cultivation with BrCa cells. Transwell® was applied to construct the co-culture system. The THP1- derived Mφ co-cultured with vector- and DRD2-transfected BrCa cells for 3 d. And primary THP-1-derived Mφ (M0) was used as control. ( D ) WB results of M1 and M2 markers after co-cultivation. ( E ) IF staining of M1 and M2 markers after co-cultivation. And THP1 cells were seeded in glass coverslips when differentiating to Mφ by PMA. M1 marker, iNOS; M2 marker, CD206. ( F ) An antibody array was used for cytokines detection of medium from BrCa cells. The medium used for detection was harvested after another 24 h incubation when finishing 3 d co-cultivation. Medium derived from BrCa cells was used as control. Fluorescence imaging (upper) and analysis of extracted data (lower) were shown. Data are presented as mean ± SD; P-value was calculated using two-tailed Student's t test. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Immunohistochemistry, Quantitative RT-PCR, Construct, Co-Culture Assay, Derivative Assay, Cell Culture, Plasmid Preparation, Transfection, Staining, Marker, Ab Array, Incubation, Fluorescence, Imaging, Two Tailed Test

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 triggers pyroptosis during crosstalk with Mφ. ( A and B ) Murinebreast cancer cell 4T1 used to construct subcutaneous tumor model. HE (A, left), IHC (A, middle and right) and TUNEL (B) assays were performed in samples derived from subcutaneous tumor model. Bars, 80 μm in (A); Bars, 75 μm in (B). ( C and D ) Pyroptosis markers were examined by qRT-PCR (C), and necroptosis, apoptosis, as well as pyroptosis markers were detected by WB (D) in vector- and DRD2-transfected BrCa cells co-cultivated with Mφ. BrCa cells cultured alone were used as controls. Data are presented as mean ± SD from biological replicates. P-value was calculated using two-tailed Student's t test. ***, p < 0.001. ns: not significant. ( E ) Markers of inflammasome (NLRP3) and pyroptosis (GSDME) were detected by WB. M1 Mφ was induced by LPS (200 ng/ml, 3 d). Medium derived from M1 Mφ was filtrated and applied to incubate Vector- and DRD2-MB231 for 3 d. Tumor cells cultured alone were used as control.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Construct, TUNEL Assay, Derivative Assay, Quantitative RT-PCR, Plasmid Preparation, Transfection, Cell Culture, Two Tailed Test

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 restricts NF-κB signaling activation by interrupting phosphorylation of TAK1. ( A and B ) Representative IF staining of DRD2 (green) and p-p65 (red) in BrCa cells treated by LPS (A) or THP1-derived Mφ (B). And BrCa cells were seeded in glass coverslips for 24h before LPS (5 μg/ml, serum-free, 24 h) or Mφ treatment. Images were taken by confocal microscopy. Vector-transfected BrCa cells were used as the control. Nuclei were stained with DAPI. Bars, 10 μm in (A); 5 μm in (B). ( C ) WB was applied to detect cytoplasmic and nuclear expression of p-p65. PCNA and Actin were used to prove the protein integrity of nucleus and cytoplasm respectively. ( D and E ) WB was used to analyze the activation status of NF-κB signaling and its upstream regulator TAK1 after being treated by LPS (5 μg/ml, serum-free, 24 h) (D) and THP1-derived Mφ (3 d) (E). ( F and G ) IB was used to confirm the binding of proteins obtained by Co-IP in MDA-MB231. Cells were treated with LPS (5 μg/ml, serum-free, 24 h) or CM (3 d) before Co-IP. The binding of DRD2, β-arrestin2 and p-p65 were analyzed by IB in DRD2-expressed MDA-MB231 with or without LPS treatment (F). The binding of DRD2 and β-arrestin2 were analyzed by IB in in Vector- and DRD2-expressed MDA-MB231 with or without CM treatment (G). ( H ) IB was sued to determine the binding of TAB1, TAK1, and β-arrestin2 in Vector- and DRD2-expressed MDA-MB231. Co-IP was used to obtain possible binding proteins of TAB1 in samples with or without LPS treatment. IgG was used as negative control, and the input protein was used as positive control. And GAPDH was used for protein integrity. CM, Conditioned Medium.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Activation Assay, Staining, Derivative Assay, Confocal Microscopy, Plasmid Preparation, Transfection, Expressing, Binding Assay, Co-Immunoprecipitation Assay, Negative Control, Positive Control

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: DRD2 antagonizes NF-κB signaling through interacting and downregulating DDX5 and eEF1A2. ( A ) qRT-PCR was used to detect mRNA expression of p65 and IL-10 in Vector- and DRD2- transfected BrCa cells. ( B ) Venn diagram showed potential binding proteins of DRD2 identified by mass spectrum analysis. And mass spectrum was performed in Co-IP isolated proteins in both MDA-MB231 and BT549. ( C ) mRNA expression of DDX5 , eEF1A2 , and ICAM-1 were analyzed by qRT-PCR in Vector- and DRD2- transfected BrCa cells. ( D ) Protein expression of DDX5 and eEF1A2 were determined by WB in Vector- and DRD2- transfected BrCa cells. The expression was analyzed in both MDA-MB231 and BT549. ( E ) The binding of DRD2, DDX5 and eEF1A2 was examined by Co-IP and IB in 293T and MDA-MB231. ( F ) ERBB1 (EGFR) and ERBB2 (HER2) mRNA expression were determined by qRT-PCR in Vector- and DRD2- transfected BrCa cells. ( G ) WB was used to detect the effect of DDX5 and eEF1A2 on NF-κB activation was analyzed by WB in MDA-MB231 by stimulation of LPS. GAPDH was used for protein integrity. Data are presented as mean ± SD from biological replicates. P-value was calculated using two-tailed Student's t test. *, p < 0.05 **, p < 0.01; ***, p < 0.001.

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Quantitative RT-PCR, Expressing, Plasmid Preparation, Transfection, Binding Assay, Co-Immunoprecipitation Assay, Isolation, Activation Assay, Two Tailed Test

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: List of antibodies used in this study

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques:

Journal: Theranostics

Article Title: Tumor suppressor DRD2 facilitates M1 macrophages and restricts NF-κB signaling to trigger pyroptosis in breast cancer

doi: 10.7150/thno.58322

Figure Lengend Snippet: Relationship between clinicopathological features and DRD2 expression in breast cancer (TCGA)

Article Snippet: Quinpirole (2μM, Cas #: 85798-08-9, Sigma-Aldrich) was applied to selectively activated DRD2. si-RNA used for knocking down DRD2 was purchased from Origene according to its Application Guide.

Techniques: Expressing