Journal: Molecular Brain

Article Title: Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo ; characterisation, subcellular localisation and modulation of the receptors

doi: 10.1186/s13041-015-0138-6

Figure Lengend Snippet: Schematic of the insulin/IGF1R signalling pathway. A simplified representation of the insulin/IGF1 signalling pathway depicting the downstream activation of Akt and p44-42 MAPK through binding of insulin or IGF1 to their respective receptors. Furthermore insulin can bind to IGF1R and IGF1 to IR and IR α-subunits/ β-subunits can form heterodimers with IGF1R α-subunits /β-subunits adding further complexity at the level of the receptors. There are also numerous downstream feedback loops within this and other signalling pathways which act to regulate signalling through this pathway at multiple levels

Article Snippet: To impair signalling through IGF1R cultures were treated with 11 μg/ml IGF1 receptor monoclonal antibody (24 h) (MAB391, R&D Systems, MN, USA).

Techniques: Activation Assay, Binding Assay

Journal: Molecular Brain

Article Title: Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo ; characterisation, subcellular localisation and modulation of the receptors

doi: 10.1186/s13041-015-0138-6

Figure Lengend Snippet: Insulin/IGF1 pathway characterisation in astrocytes. Immunoblots of astrocytes cultured in either serum containing or serum deprived media with additional supplementation from either a recombinant human 1 μM insulin or b 11.2nM recombinant human IGF1. Respresentative images from blots probed with antibodies against IRβ, IGF1Rβ, IRS1, IRS2, pAkt, Total Akt, p44/42 MAPK are shown. *α-tubulin was used as a loading control for blots and a representative loading control is shown. Molecular weight markers are indicated (kDa)

Article Snippet: To impair signalling through IGF1R cultures were treated with 11 μg/ml IGF1 receptor monoclonal antibody (24 h) (MAB391, R&D Systems, MN, USA).

Techniques: Western Blot, Cell Culture, Recombinant, Control, Molecular Weight

Journal: Molecular Brain

Article Title: Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo ; characterisation, subcellular localisation and modulation of the receptors

doi: 10.1186/s13041-015-0138-6

Figure Lengend Snippet: Impairment of IGF1 signalling using a monoclonal IGF1 antibody (MAB391). Human astrocytes treated with 11 μg/ml MAB391 for 24 h ( a ). Representative immunoblots demonstrate reductions in IGF1Rβ, IRβ and pAkt in response to MAB391 with no impact on total IRS1 or downstream signalling through p44/42 MAPK. *α-tubulin was used as a loading control for blots and a representative loading control is shown. Molecular weight markers are indicated (kDa). b Bar charts show quantification of immunoblots, pAkt was normalised to Akt/α-tubulin. c Immunofluorescence showing the reduction in IGF1R, scale bars represent 10 μM. d qPCR analysis of IGF1Rβ RNA after 24 h show no differences at the RNA level. Data represents mean + SEM ( n = 3, 3 replicates/experiments, Unpaired t -test, ** p < 0.01)

Article Snippet: To impair signalling through IGF1R cultures were treated with 11 μg/ml IGF1 receptor monoclonal antibody (24 h) (MAB391, R&D Systems, MN, USA).

Techniques: Western Blot, Control, Molecular Weight, Immunofluorescence