drd3 polyclonal antibody, cy5 conjugated  (Bioss)

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: The expression and distribution of dopamine receptors, D1R and D3R in mouse hearts: (A) Representative photomicrographs showing immunofluorescence staining of D1R and D3R which co-localized with vimentin, a cardiac fibroblast-specific marker, in the heart tissue sections. No primary antibody control sections were incubated with the antibody diluent alone and no primary antibody, followed by incubation with secondary antibodies and DAPI (the nuclear stain), which confirms the antibody specificity of D1R and D3R. (B) Gene expression was determined using TaqMan primers by real-time qPCR. The normalized -dCT values indicate that there is relatively higher expression of D1R compared to D3R in cardiac tissue. (C) Receptor protein expression determined by western blot using receptor specific antibodies as described in methods section.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Marker, Incubation, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: The expression and distribution of dopamine receptors, D1R and D3R in mouse primary cardiac fibroblasts: (A) Representative photomicrographs showing immunofluorescence staining of D1R and D3R which co-localized with cardiac fibroblasts specific marker vimentin staining in the isolated WT and D3KO primary cardiac fibroblasts in culture. Cardiac fibroblast expression of D1R and D3R is not lost when primary cells are maintained in cell culture. As expected, there is no observable staining for D3R in D3KO cardiac fibroblasts. No primary antibody control images show the specificity of the primary antibodies used. (B) Gene expression determined using TaqMan primers by real-time qPCR. The normalized -dCT values indicate that there is relatively higher expression D1R compared to D3R in cardiac fibroblasts. (C) Receptor protein expression determined by western blot using receptor specific antibodies as described in methods section.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Marker, Isolation, Cell Culture, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Loss of Function in Dopamine D3 Receptor Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation in vitro

doi: 10.3389/fcvm.2021.732282

Figure Lengend Snippet: Cardiac fibroblast cell proliferation and migration studies: (A) Cardiac fibroblast cell proliferation between WT + vehicle (black circle), WT treated with SB (closed blue square), and D3KO + vehicle (red circle) over a time-period of 36 h. Cells were incubated in DMEM/F-12 growth media supplemented with 10% FBS; n = 3 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean. (B) Cardiac fibroblast cell migration distance in response to a scratch injury over a 24-h time-period. Migration distance in μm between WT + vehicle (black circle), D3KO + vehicle (red circle), and WT cells treated with D3R specific agonist, pramipexole (blue circle). Cells were incubated in DMEM/F-12 growth media supplemented with 0.5% FBS; n = 9 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean. Statistical significance between WT treatment and D3KO + vehicle. (C) Cardiac fibroblast cell migration distance in response to a scratch injury over a 24-h time-period. Migration distance in μm of D3KO + vehicle cells (red circle), D3KO cells treated with pramipexole (closed blue square), and D3KO cell treated with SB (green square), compared against WT + vehicle cells (black circle); n = 9 for each treatment at each time-point, * p < 0.05 statistically significant. Bars represent standard error of mean.

Article Snippet: Both cells and tissues were then incubated with primary antibodies for Vimentin [M0725, lot #027(102), Dako, 1:500 dilution], D1R (NB110-60017AF488, lot #B-3-101620, Novus Biologics, 1:500 dilution), and D3R (bs-1743R-Cy5, lot #AF12125641, Bioss, 1:500 dilution) overnight at 4°C.

Techniques: Migration, Incubation