Journal: Cells

Article Title: Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds

doi: 10.3390/cells11020247

Figure Lengend Snippet: ( a ) Immunofluorescence of specific neuronal markers (β-III tubulin, MAP2 and NeuN) in undifferentiated cells (SH-SY5Y cells) and ( b ) differentiated cells (sensory neuron-like cells). Representative images of five areas. Negative control of immunostaining. Scale bar: 50 µm. ( c ) Relative quantification of immunostaining for βIII-tubulin, MAP2, NeuN representing the ratio expression in the sensory neuron-like cells and SH-SY5Y cells. The data were analyzed by t -test, * p < 0.05; ** p < 0.001). The data represent three independent and similar assays.

Article Snippet: Cells were incubated with primary antibodies diluted in PBS and 3% BSA: chicken anti-neuron specific β-III tubulin diluted 1:500 (Merck Millipore, Burlington, MA, USA) and rabbit anti-RAGE diluted 1:200 (Abcam, Waltham, MA, USA); or incubated with chicken anti-neuron specific β-III tubulin diluted 1:500 and rabbit anti-Galectin-3 (1:500, Abcam, Waltham, MA, USA) overnight at 4 °C.

Techniques: Immunofluorescence, Negative Control, Immunostaining, Expressing

Journal: Cells

Article Title: Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds

doi: 10.3390/cells11020247

Figure Lengend Snippet: ( a ) Representative images of immunofluorescence of RAGE expression. Neuronal class β-III tubulin (red) 1:500; Hoechst-33342 was used to stain nuclei (blue) 1:3000; RAGE receptor (green) 1:200 in control cells (differentiated/ without ECM) and cells treated with ECM-NC, ECM-GC, ECM-NC IL-1β. Negative control of immunostaining. Magnification 40×. Scale bar: 25 µm. ( b ) Relative quantification of immunostaining for RAGE representing the ratio between the experimental and the control group. The data were analyzed by t -test. The data represent three independent and similar assays. Data were analyzed by one-way ANOVA with post hoc testing by Dunnett p > 0.05).

Article Snippet: Cells were incubated with primary antibodies diluted in PBS and 3% BSA: chicken anti-neuron specific β-III tubulin diluted 1:500 (Merck Millipore, Burlington, MA, USA) and rabbit anti-RAGE diluted 1:200 (Abcam, Waltham, MA, USA); or incubated with chicken anti-neuron specific β-III tubulin diluted 1:500 and rabbit anti-Galectin-3 (1:500, Abcam, Waltham, MA, USA) overnight at 4 °C.

Techniques: Immunofluorescence, Expressing, Staining, Negative Control, Immunostaining

Journal: Cells

Article Title: Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds

doi: 10.3390/cells11020247

Figure Lengend Snippet: ( a ) Representative images of immunofluorescence of Galectin-3 expression. Neuronal class β-III tubulin (red) 1:500; Hoechst-33342 was used to stain nuclei (blue) 1:3000; Galectin-3 receptor (green) 1:200 in control cells (differentiated/ without ECM) and cells treated with ECM-NC, ECM-GC, ECM-NC IL-1β. Negative control of immunostaining. Magnification 40×. Scale bar: 25 µm. ( b ) Relative quantification of immunostaining for Galectin-3 representing the ratio between the experimental and the control group. The data were analyzed by t -test. The data represent three independent and similar assays. Data were analyzed by one-way ANOVA with post hoc testing by Dunnett p > 0.05).

Article Snippet: Cells were incubated with primary antibodies diluted in PBS and 3% BSA: chicken anti-neuron specific β-III tubulin diluted 1:500 (Merck Millipore, Burlington, MA, USA) and rabbit anti-RAGE diluted 1:200 (Abcam, Waltham, MA, USA); or incubated with chicken anti-neuron specific β-III tubulin diluted 1:500 and rabbit anti-Galectin-3 (1:500, Abcam, Waltham, MA, USA) overnight at 4 °C.

Techniques: Immunofluorescence, Expressing, Staining, Negative Control, Immunostaining

Journal: Cells

Article Title: Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds

doi: 10.3390/cells11020247

Figure Lengend Snippet: ( a ) Representative images of immunofluorescence show neurite length in sensory neuron-like cells. Arrows represent the reduced neuron interconnection and retraction of neurons outgrowth. ( b ) Quantification of neurite length in neuron-like cells based on β-III tubulin immunostaining. Data from 72 h of incubation. Magnification 20×. Scale bar: 100 µm. Data were analyzed by t -test (* p < 0.05 different from control), ECM-NC (normal collagen), ECM-GC (glycated collagen). Three independent and similar assays were performed in triplicate.

Article Snippet: Cells were incubated with primary antibodies diluted in PBS and 3% BSA: chicken anti-neuron specific β-III tubulin diluted 1:500 (Merck Millipore, Burlington, MA, USA) and rabbit anti-RAGE diluted 1:200 (Abcam, Waltham, MA, USA); or incubated with chicken anti-neuron specific β-III tubulin diluted 1:500 and rabbit anti-Galectin-3 (1:500, Abcam, Waltham, MA, USA) overnight at 4 °C.

Techniques: Immunofluorescence, Immunostaining, Incubation