Table 2.
Viruses, antibodies, reagents, and incubation parameters.
| Name | Source | Incubation | Experiment |
|---|---|---|---|
| AAV-hSyn-DIO-hM3D(Gq)-mCherry | Addgene, #44361-AAV8 | Chemogenetic stimulation for feeding behavior, cFos | |
| AAV8-hSyn-DIO-mCherry | Addgene #50459-AAV8 | Control virus for chemogenetic stimulation, cFos | |
| Alexa Fluor® 488 Donkey Anti-Rabbit IgG | Jackson ImmunoResearch, 711-545-152 | 1:500, 3 h, RT | Immunofluorescent labeling |
| Alexa Fluor® 647 Donkey Anti-Rabbit IgG | Jackson ImmunoResearch, 711-605-152 | 1:500, 3 h, RT | Immunofluorescent labeling |
| Biotin-SP Donkey Anti-Rabbit IgG | Jackson ImmunoResearch, 711-065-152 | 1:1 K, 2 h, RT | Immunoperoxidase labeling |
| Chicken Anti-Green Fluorescent Protein | Abcam, ab13970, RRID:AB_300798 | 1:1 K, 24 h, RT | Immunofluorescent labeling |
| Cy™3 Donkey Anti-Guinea Pig IgG (H + L) | Jackson ImmunoResearch, 706-165-148 | 1:300, 40 h, 37 °C | CLARITY |
| 1:500, 3 h, RT | Immunofluorescent labeling | ||
| Donkey anti-Rabbit IgG, HRP conjugated | Millipore Sigma, AP 182 P | 1:500, 40 h, 37 °C | CLARITY |
| 1:500, 3 h, RT | TSAP-enhanced Immunofluorescent labeling | ||
| Guinea Pig Anti-Red Fluorescent Protein | Synaptic Systems, 390 004, RRID:AB_2737052 | 1:15 K, 48 h, RT | Immunofluorescent labeling |
| 1:10 K, 24 h, RT | Immunoperoxidase labeling | ||
| iCre mRNA | Advanced Cell Diagnostics, 423321-C2, Accession No. AY056050.1 | 1:50, 2 h 40 °C | RNAscope labeling |
| Mineral oil | VWR Life Science, J217 | 100%, 3 h, 37 °C | CLARITY |
| Mouse Anti-Dopamine β Hydroxylase | Millipore Sigma, MAB308, RRID:AB_2245740 | 1:8 K, 48 h, RT | Immunofluorescent labeling |
| Rabbit Anti c-Fos (9F6) | Cell Signaling, #2250, RRID:AB_2247211 | 1:10 K, 24 h, RT | Immunoperoxidase labeling |
| Rabbit Anti-Glucagon-Like Peptide 1 (7–37) | Peninsula Laboratories, T-4363, RRID:AB_518978 | 1:10 K, 90 h, 37 °C | CLARITY |
| 1:25 K, 48 h, RT | TSAP-enhanced Immunofluorescent labeling | ||
| Rn-Gcg mRNA | Advanced Cell Diagnostics, 315471-C2, Accession No. NM_012707.2 | 1:50, 2 h 40 °C | RNAscope labeling |
| TSA™Plus Cyanine 5 | AKOYA Biosciences, SAT705A001KT | Clarity: 1:400, 40 h, 37 °C | CLARITY |
| 1:1 K & 1:2 K, 30min, 40 °C | RNAscope labeling | ||
| 2,20-Thiodiethanol | Sigma-Aldrich, 166782 | 15%, 30%, 45%, 60%, 5 h each, RT | CLARITY |
| 2% bis-Acrylamide | RPI Research Products, A11275 | CLARITY | |
| 40% Acrylamide Solution | RPI Research Products, A11265 | CLARITY |