Fig 1: Total protein separated by SDS-PAGE, Resolved on 12% resolving gel, stained with Commassie blueNotes: A total of 10 wells of gel was loaded with 9 samples and protein marker (M) of known molecular weight to check the presence of protein and their resolution. The representative images of the protein blot of different areas of the brain and β-actin, for GLP-1 expression. At the bottom, a graphical representation of the GLP-1 protein expression is seen. Densitometric analysis was done using the Image J software by selecting bandwidth and measurement of intensity. The quantification of the bands was then performed using β-actin as a control. Each of the brain areas studied is shown on the X axis and the ratio of band intensities of the sample (from each brain area)/β-actin is shown on the Y axis.
Fig 2: In the upper panel, melt curve analysis is shown-performed at the end of the PCR cycle, to confirm the specificity of primer annealingIn the lower panel, a graphical representation of the GLP-1 recep areas of the brain is shown. The Y axis represents the mean of delta cycle threshold values for these areas.
Fig 3: Microscopic anatomy of various cerebral cortical areas after immunostaining using GLP-1 monoclonal antibodyNotes: The macroscopic view of the cerebrum in the mid-sagittal section depicts the prefrontal (PF), the frontal (F), and the parietal (P) cortical areas. Below that, an inferior view of the cerebrum, dissected to demonstrate the hippocampus is shown in the left lower corner. The parietal cortex at 40x magnification depicts moderate immune staining in the neuropil and scattered mild staining in neurons. The frontal cortex at 40x magnification depicts cytoplasmic staining in large pyramidal cells of cortical layer 5. The prefrontal cortex shows moderate immunoreactivity in the granule cells and surrounding neuropil, including a vessel wall. The hippocampal trilaminar cortex with scattered immunoreactivity is observed at 4x magnification. At 40x magnification, moderate patchy IR is seen in large pyramidal cells and few fibers in the stratum pyramidal of the Hippocampus.
Fig 4: The upper panel depicting macroscopic mid sagittal view of the thalamus and the hypothalamusNotes: The diagrammatic representation of the thalamic medial dorsal nucleus (MDN) and the hypothalamic paraventricular nucleus (PVN) is shown in the section. The thalamic MDN at 20x magnification depicts moderated immunoreactivity in the fibers and a few large neurons. The hypothalamic PVN shows magnocellular neurons in the background of small neurons at 40x. immunoreactivity is seen in the magnocellular neurons as cytoplasmic stippling forming a crescent. Small neurons did not take the stain. The lower panel depicts a macroscopic view of the olive in the medulla, followed by the microscopic view of the inferior olivary nucleus. A typical crumpled bag appearance of the inferior olivary nucleus can be seen at 10x. In the inset, the pattern of staining in the neuron can be appreciated in the form of cytoplasmic punctate moderate intensity immunoreactivity, at 40x magnification. The pons at 40 x magnification shows a group of pontine nuclei and surrounding neuropil with moderate immunoreactivity. Immunostaining has been done using GLP-1 monoclonal antibody.
Supplier Page from Abcam for Anti-GLP-1 antibody [1B7B4]