Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Cell adhesion molecule close homolog of L1 binds to the dopamine receptor D2 and inhibits the internalization of its short isoform.

doi: 10.1096/fj.201900577RRRR

Figure Lengend Snippet: FIGURE 9 Interaction of CHL1 and DRD2 on TH- and DARP32-positive neurons in striatal sections. Proximity ligation assay using goat anti-CHL1 and mouse anti-DRD2 antibodies was combined with immunostaining using rabbit anti-DARPP32 (A) or anti-TH (B) antibodies to analyze tissue sections from 12- to 18-week-old CHL1+/+ mice. Nuclei are stained with DAPI (blue). Representative images are shown. Close- ups of two regions (without DAPI staining) are indicated by boxes and arrowheads indicate red spots indicating close molecular interaction of CHL1 with DRD2. Scale bars: 10 µm. Three independent experiments were performed with different sets of animals

Article Snippet: The polyclonal rabbit antibodies against TH phosphorylated at Ser40 (pSer-TH) (Bio-Rad/AbD Serotec Cat# AHP912, RRID:AB_567401) and against DARPP32 phosphorylated at Thr34 (pThr34-DARPP32) (Bio-Rad/AbD Serotec Cat# AHP897, RRID:AB_566944) were from BioRad (Puchheim, Germany).

Techniques: Proximity Ligation Assay, Immunostaining, Staining

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Cell adhesion molecule close homolog of L1 binds to the dopamine receptor D2 and inhibits the internalization of its short isoform.

doi: 10.1096/fj.201900577RRRR

Figure Lengend Snippet: FIGURE 10 Interaction of CHL1 and DRD2 on TH- and DARP32-positive cells in cultures of ventral midbrain and striatum. Cultures of ventral midbrain (A) or striatum (B) were analyzed by proximity ligation assay using goat anti-CHL1 and mouse anti-DRD2 antibodies combined with immunofluorescent staining using rabbit anti-TH or anti-DARPP32 antibodies. Nuclei are stained with DAPI (blue). Representative images of different cells are shown. Red spots indicate close molecular interaction between CHL1 and DRD2. Scale bars: 10 µm. Three independent experiments were performed with different sets of animals

Article Snippet: The polyclonal rabbit antibodies against TH phosphorylated at Ser40 (pSer-TH) (Bio-Rad/AbD Serotec Cat# AHP912, RRID:AB_567401) and against DARPP32 phosphorylated at Thr34 (pThr34-DARPP32) (Bio-Rad/AbD Serotec Cat# AHP897, RRID:AB_566944) were from BioRad (Puchheim, Germany).

Techniques: Proximity Ligation Assay, Staining

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Cell adhesion molecule close homolog of L1 binds to the dopamine receptor D2 and inhibits the internalization of its short isoform.

doi: 10.1096/fj.201900577RRRR

Figure Lengend Snippet: FIGURE 11 Reduced DRD2 and pSer40-TH levels in the dorsal striatum and reduced pThr34-DARPP32 levels in the ventral striatum in the absence of CHL1. The dorsal and ventral parts of the striatum were isolated from 10- to 13-week-old CHL1+/+ and CHL1−/− mice and subjected to Western blot analysis with anti-DRD2 and anti-GAPDH antibodies (A), anti-pSer40-TH and anti-TH antibodies (B) or anti-pThr34-DARPP32 and anti-DARPP32 (C) antibodies. Protein levels were determined by densitometry and DRD2 levels relative to GAPDH levels (A), pSer40-TH levels relative to total TH levels (B) and pThr34-DARPP32 levels relative to total DARPP32 levels (C) were calculated. A-C, Representative Western blots (left panels) are shown and mean values + standard error of the mean from eight CHL1−/− and 11 CHL1+/+ mice (right panels) are shown for the relative levels of DRD2 (A), pSer40-TH (B) and pThr34-DARPP32 (C) (Kruskal-Wallis test with post-hoc Dunn´s multiple comparison test; *P < .05, **P < .01). A, Lanes not adjacent to each other but derived from the same blot are separated by a vertical line

Article Snippet: The polyclonal rabbit antibodies against TH phosphorylated at Ser40 (pSer-TH) (Bio-Rad/AbD Serotec Cat# AHP912, RRID:AB_567401) and against DARPP32 phosphorylated at Thr34 (pThr34-DARPP32) (Bio-Rad/AbD Serotec Cat# AHP897, RRID:AB_566944) were from BioRad (Puchheim, Germany).

Techniques: Isolation, Western Blot, Comparison, Derivative Assay

Journal: bioRxiv

Article Title: Neuroprotective effects of hepatoma-derived growth factor in models of Huntington’s disease

doi: 10.1101/2023.02.23.529222

Figure Lengend Snippet: a , Western blots of the indicated brain regions from 8-week-old R6/2 mice and WT littermate controls. HDGF band is indicated on the right, the upper band is unspecific. Total protein detection was used as loading control. b , Quantification of HDGF protein quantity in the indicated brain regions of 8- and 12-week-old R6/2 mice, normalized first to total protein and then to the average value of the littermate controls. N=4-5 mice per group. Unpaired two-tailed t-test, per brain region and age group. No significant differences were observed. CE, cerebellum; HP, hippocampus; CX, cortex; ST, striatum. c , Images of brain sections from 12-week-old R6/2 and WT mice immunostained for HDGF. DARPP32 and neurogranin (NGRN) were used as markers of striatal MSNs and cortical PCs, respectively. Nuclei were counterstained with DAPI. d , Quantification of HDGF immunofluorescence intensity in striatal MSNs and cortical PCs of R6/2 mice and WT controls. Values were background-subtracted and normalized to the fluorescence intensity of the highest expressing cell in the field of view. N=3 mice per group. Unpaired two-tailed t-test, per cell type. No significant differences were observed. Scale bar in c, 20 µm.

Article Snippet: The following primary antibodies were used: rabbit anti-HDGF (Abcam, ab128921, 1:500), mouse anti-HTT (EM48, Chemicon, MAB5374, 1:500), mouse anti-Flag (Origene, TA50011, 1:1,000), goat anti-ChAT (Chemicon, AB144, 1:500), goat anti-DARPP32 (LifeSpan Biosciences, LS-C150127, 1:300), mouse anti-NGRN (R&D Systems, MAB7947, 1:60), chicken anti-GFAP (Origene, AP31806PU-N, 1:2,000, with antigen retrieval), mouse anti-APC (Calbiochem, OP80, 1:20, with antigen retrieval), and goat anti-IBA1 (Abcam, ab107159, 1:1,000, with antigen retrieval).

Techniques: Western Blot, Two Tailed Test, Immunofluorescence, Fluorescence, Expressing

Journal: PLoS Biology

Article Title: Absence of Ret Signaling in Mice Causes Progressive and Late Degeneration of the Nigrostriatal System

doi: 10.1371/journal.pbio.0050039

Figure Lengend Snippet: Immunohistochemical stainings of dorsal striatum of 2-y-old control (A, D, and G) and DAT-Ret lx/lx mutants (B, E, and H) for NeuN (A and B), DARPP-32 (D and E), or parvalbumin (G and H). Histograms showing the number of NeuN-positive (C), DARPP-32–positive (F), and parvalbumin-positive cells (I) in DAT-Ret lx/lx mutants and age-matched controls ( n = 3–5 each genotype). Note also the reduced staining intensities for NeuN and DARPP-32 in DAT-Ret lx/lx compared to control mice. *, p < 0.05; **, p < 0.01 (Student t -test). Scale bars indicate 50 μm.

Article Snippet: Primary antibodies used were goat anti-Ret (1:25; RDI, Flanders, New Jersey, United States, or Neuromics, North Field, Minnesota, United States), monoclonal mouse anti-tyrosine hydroxylase (1:2,000; DiaSorin, Stillwater, Massachusetts, United States), rabbit anti-dopa decarboxylase (1:100; Chemicon/Millipore, Billerica, Massachusetts, United States), monoclonal mouse anti–β-galactosidase (1:50; Sigma, St. Louis, Missouri, United States), rat anti-dopamine transporter (1:500; Chemicon/Millipore), rabbit anti-Pitx3 (1:1,000, provided by M. P. Smidt [ ]), monoclonal mouse anti-NeuN (1:200; Chemicon/Millipore), rabbit anti-GFAP (1:500; DakoCytomation, Glostrup, Denmark), rabbit anti–DARPP-32 (1:50; United States Biological, Swampscott, Massachusetts, United States), monoclonal mouse anti-parvalbumin (1:10,000; Swant, Bellinzona, Switzerland), rabbit anti–Iba-1 (1:1,000; Wako, Neuss, Germany), monoclonal rat anti-MAC1 (1:200; Serotec, Kidlington, United Kingdom).

Techniques: Immunohistochemical staining, Staining

Journal: British Journal of Pharmacology

Article Title: Prenatal and postnatal alcohol exposure increases vulnerability to cocaine addiction in adult mice

doi: 10.1111/bph.14901

Figure Lengend Snippet: Primary antibodies

Article Snippet: The Immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 ). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Primary antibody Description Host Dilution Company Catalogue number RRID ΔFosB Delta FosB Mouse 1:250 Abcam #ab11959 AB_298732 CREB cAMP‐response element binding protein Rabbit 1:500 MERCK #04‐767 AB_1586959 D 1 R Dopamine receptor 1 Rabbit 1:500 Abcam #ab20066 AB_445306 D 2 R Dopamine receptor 2 Rabbit 1:500 MERCK #324393 AB_10681601 DARPP‐32 Dopamine‐ and cAMP‐regulated phosphoprotein Rabbit 1:1,000 Cell Signaling #2306 AB_823479 GAPDH Glyceraldehyde‐3‐phosphate dehydrogenase Mouse 1:2,500 Santa Cruz Biotechnology #sc‐365062 AB_10847862 GluA1 AMPA receptor subunit 1 Rabbit 1:1,000 MERCK #ABN241 AB_2721164 GluA2 AMPA receptor subunit 2 Rabbit 1:1,000 MERCK #AB1768‐I AB_2313802 pCREB Phosphorylated CREB Rabbit 1:1,000 MERCK #06‐519 AB_310153 pDARPP‐32 (Thr34) Phosphorylated DARPP‐32 (Thr34) Rabbit 1:1,000 MERCK #ab9206 AB_347689 β‐Tubulin β‐Tubulin Mouse 1:5,000 BD PharMingen #556321 AB_396360 β‐Tubulin Class III β‐tubulin Rabbit 1:1,000 Abcam #ab6046 AB_2210370 Open in a separate window Primary antibodies 2.7.

Techniques: Binding Assay

Journal: British Journal of Pharmacology

Article Title: Prenatal and postnatal alcohol exposure increases vulnerability to cocaine addiction in adult mice

doi: 10.1111/bph.14901

Figure Lengend Snippet: GluA1/GluA2 ratio and striatal ΔFosB expression were altered in PLAE mice after cocaine‐primed reinstatement session. (a–f) Western blot analyses of GluA1/GluA2, pCREB/CREB, pDARPP‐32/DARPP‐32, ΔFosB, D1R, and D2R protein expression in the PFC and STR of PLAE and control mice following cocaine‐induced reinstatement. Student's t test, * P < .05 control versus PLAE group (n = 5–6 per group). The numbers in the bars represent the amount of individuals in the group. The lower panels show representative fluorescent immunoblots

Article Snippet: The Immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 ). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Primary antibody Description Host Dilution Company Catalogue number RRID ΔFosB Delta FosB Mouse 1:250 Abcam #ab11959 AB_298732 CREB cAMP‐response element binding protein Rabbit 1:500 MERCK #04‐767 AB_1586959 D 1 R Dopamine receptor 1 Rabbit 1:500 Abcam #ab20066 AB_445306 D 2 R Dopamine receptor 2 Rabbit 1:500 MERCK #324393 AB_10681601 DARPP‐32 Dopamine‐ and cAMP‐regulated phosphoprotein Rabbit 1:1,000 Cell Signaling #2306 AB_823479 GAPDH Glyceraldehyde‐3‐phosphate dehydrogenase Mouse 1:2,500 Santa Cruz Biotechnology #sc‐365062 AB_10847862 GluA1 AMPA receptor subunit 1 Rabbit 1:1,000 MERCK #ABN241 AB_2721164 GluA2 AMPA receptor subunit 2 Rabbit 1:1,000 MERCK #AB1768‐I AB_2313802 pCREB Phosphorylated CREB Rabbit 1:1,000 MERCK #06‐519 AB_310153 pDARPP‐32 (Thr34) Phosphorylated DARPP‐32 (Thr34) Rabbit 1:1,000 MERCK #ab9206 AB_347689 β‐Tubulin β‐Tubulin Mouse 1:5,000 BD PharMingen #556321 AB_396360 β‐Tubulin Class III β‐tubulin Rabbit 1:1,000 Abcam #ab6046 AB_2210370 Open in a separate window Primary antibodies 2.7.

Techniques: Expressing, Western Blot