Journal: Neurology International

Article Title: Hyperglycaemia Aggravates Oxidised Low-Density Lipoprotein-Induced Schwann Cell Death via Hyperactivation of Toll-like Receptor 4

doi: 10.3390/neurolint16020027

Figure Lengend Snippet: Primers used for quantitative real-time PCR.

Article Snippet: For immunocytochemistry, the cells were seeded onto coverslips and cultured for 24 h (followed by fixation with 4% paraformaldehyde for 10 min at 25 °C), blocked with 1% bovine serum albumin containing 0.1% tween-20, incubated with anti-TLR4 primary antibody (1:200) for 1 h at 25 °C, washed with PBS, incubated with Alexa 488-conjugated secondary antibody (1:500) for 1 h at 25 °C, and mounted with Fluoroshield and DAPI (ImmunoBioScience Corp., Mukilteo, WA, USA).

Techniques:

Journal: Neurology International

Article Title: Hyperglycaemia Aggravates Oxidised Low-Density Lipoprotein-Induced Schwann Cell Death via Hyperactivation of Toll-like Receptor 4

doi: 10.3390/neurolint16020027

Figure Lengend Snippet: Expression and localisation of TLR4 in IMS32 cells. ( A ) TLR4 mRNA expression in IMS32 cells. ( B ) IMS32 cells were grown on coverslips for 24 h. The cells were fixed with 4% paraformaldehyde, permeabilised, and immunostained with anti-TLR4 antibody (green). The nuclei were stained with DAPI (blue). DIC, differential interference contrast scale bar, 5 μm.

Article Snippet: For immunocytochemistry, the cells were seeded onto coverslips and cultured for 24 h (followed by fixation with 4% paraformaldehyde for 10 min at 25 °C), blocked with 1% bovine serum albumin containing 0.1% tween-20, incubated with anti-TLR4 primary antibody (1:200) for 1 h at 25 °C, washed with PBS, incubated with Alexa 488-conjugated secondary antibody (1:500) for 1 h at 25 °C, and mounted with Fluoroshield and DAPI (ImmunoBioScience Corp., Mukilteo, WA, USA).

Techniques: Expressing, Staining

Journal: Neurology International

Article Title: Hyperglycaemia Aggravates Oxidised Low-Density Lipoprotein-Induced Schwann Cell Death via Hyperactivation of Toll-like Receptor 4

doi: 10.3390/neurolint16020027

Figure Lengend Snippet: Hyperglycaemia and oxLDL synergistically upregulate TLR4 expression in IMS32 cells. IMS32 cells were cultured in either normo- or hyperglycaemic conditions in the presence or absence of oxLDL (150 μg/mL) for 24 h. ( A ) Relative mRNA expressions of TLR4 were tested by RT-qPCR ( n = 6 per group). ( B ) Immunoblot analysis of TLR4 expression ( n = 12 per group). Data are presented as the means of two ( A ) or three independent experiments. * p < 0.05, ** p < 0.01. IMS32, immortalised mouse Schwann; oxLDL, oxidised low-density lipoprotein; RT-qPCR, quantitative reverse transcription polymerase chain reaction; TLR4, toll-like receptor 4.

Article Snippet: For immunocytochemistry, the cells were seeded onto coverslips and cultured for 24 h (followed by fixation with 4% paraformaldehyde for 10 min at 25 °C), blocked with 1% bovine serum albumin containing 0.1% tween-20, incubated with anti-TLR4 primary antibody (1:200) for 1 h at 25 °C, washed with PBS, incubated with Alexa 488-conjugated secondary antibody (1:500) for 1 h at 25 °C, and mounted with Fluoroshield and DAPI (ImmunoBioScience Corp., Mukilteo, WA, USA).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Western Blot, Reverse Transcription, Polymerase Chain Reaction

Journal: Neurology International

Article Title: Hyperglycaemia Aggravates Oxidised Low-Density Lipoprotein-Induced Schwann Cell Death via Hyperactivation of Toll-like Receptor 4

doi: 10.3390/neurolint16020027

Figure Lengend Snippet: TLR4 inhibition attenuated the hyperglycaemia and oxLDL-mediated cell death. IMS32 cells were pre-treated with or without TAK-242 (100 nM) for 2 h and cultured in normo- or hyperglycaemic conditions in the presence or absence of oxLDL (150 μg/mL) for 24 h. Cell viability was quantified by the MTT assay ( n = 5 per group). Data are representative of at least three independent experiments. * p < 0.05, ** p < 0.01, comparisons between indicated groups. IMS32, immortalised mouse Schwann; MTT, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; n.s., not significant; oxLDL, oxidised low-density lipoprotein; TLR4, toll-like receptor 4.

Article Snippet: For immunocytochemistry, the cells were seeded onto coverslips and cultured for 24 h (followed by fixation with 4% paraformaldehyde for 10 min at 25 °C), blocked with 1% bovine serum albumin containing 0.1% tween-20, incubated with anti-TLR4 primary antibody (1:200) for 1 h at 25 °C, washed with PBS, incubated with Alexa 488-conjugated secondary antibody (1:500) for 1 h at 25 °C, and mounted with Fluoroshield and DAPI (ImmunoBioScience Corp., Mukilteo, WA, USA).

Techniques: Inhibition, Cell Culture, MTT Assay

Journal: Neurology International

Article Title: Hyperglycaemia Aggravates Oxidised Low-Density Lipoprotein-Induced Schwann Cell Death via Hyperactivation of Toll-like Receptor 4

doi: 10.3390/neurolint16020027

Figure Lengend Snippet: TLR4 inhibition suppressed caspase-3-dependent apoptosis. IMS32 cells were pre-treated with or without TAK-242 (100 nM) for 2 h and cultured in either normo- or hyperglycaemic conditions in the presence or absence of oxLDL (150 μg/mL) for 3 h. Caspase-3 activity was measured. * p < 0.05, ** p < 0.01, comparisons between indicated groups. n.s., not significant; IMS32, immortalised mouse Schwann; TLR4, toll-like receptor 4.

Article Snippet: For immunocytochemistry, the cells were seeded onto coverslips and cultured for 24 h (followed by fixation with 4% paraformaldehyde for 10 min at 25 °C), blocked with 1% bovine serum albumin containing 0.1% tween-20, incubated with anti-TLR4 primary antibody (1:200) for 1 h at 25 °C, washed with PBS, incubated with Alexa 488-conjugated secondary antibody (1:500) for 1 h at 25 °C, and mounted with Fluoroshield and DAPI (ImmunoBioScience Corp., Mukilteo, WA, USA).

Techniques: Inhibition, Cell Culture, Activity Assay