Journal: Journal of Cell Science

Article Title: LDL receptor-related protein-1 is a sialic-acid-independent receptor for myelin-associated glycoprotein that functions in neurite outgrowth inhibition by MAG and CNS myelin

doi: 10.1242/jcs.113191

Figure Lengend Snippet: LRP1 antagonism or gene-silencing attenuates the effects of MAG-CHO cells on neurite outgrowth in PC12 and N2a cells. (A) PC12 and N2a cells were plated on R2-CHO or MAG-CHO cells and cultured for 48 h in the presence of RAP or GST (200 nM). Neurite outgrowth was determined. Results were normalized against those obtained when cells were plated on R2-CHO cells with GST (means±s.e.m., n = 3, **P<0.01). (B) PC12 and N2a cells were transfected with NTC or LRP1-specific siRNA (siLRP1) prior to plating on R2-CHO or MAG-CHO cells for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, **P<0.01). Scale bars: 100 µm.

Article Snippet: For expression of recombinant proteins, transfected CHO-K1 cells were cultured in Power-CHO CD medium (Lonza, Anaheim, CA).

Techniques: Cell Culture, Transfection

Journal: Journal of Cell Science

Article Title: LDL receptor-related protein-1 is a sialic-acid-independent receptor for myelin-associated glycoprotein that functions in neurite outgrowth inhibition by MAG and CNS myelin

doi: 10.1242/jcs.113191

Figure Lengend Snippet: LRP1 inactivation attenuates the effects of MAG-CHO cells on neurite outgrowth in CGNs. (A) CGNs pre-treated with RAP or GST were plated on MAG-CHO or R2-CHO cells and cultured for 48 h. Neurite outgrowth was measured (means±s.e.m., n = 3, **P<0.01). (B) CGNs transfected with NTC or LRP1-specific siRNA (siLRP1) were plated on MAG-CHO or R2-CHO cells and cultured for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, *P<0.05, **P<0.01). (C) Mouse CGNs, in which both LRP1 genes were floxed, were infected with HSV1-Cre or control HSV1 and cultured for 24 h. LRP1 expression was determined by immunoblot analysis. Neurite outgrowth on MAG-CHO and R2-CHO cells was determined (means±s.e.m., n = 3, **P<0.01). Scale bars: 100 µm.

Article Snippet: For expression of recombinant proteins, transfected CHO-K1 cells were cultured in Power-CHO CD medium (Lonza, Anaheim, CA).

Techniques: Cell Culture, Transfection, Infection, Expressing, Western Blot

Journal: Journal of Cell Science

Article Title: LDL receptor-related protein-1 is a sialic-acid-independent receptor for myelin-associated glycoprotein that functions in neurite outgrowth inhibition by MAG and CNS myelin

doi: 10.1242/jcs.113191

Figure Lengend Snippet: Soluble LRP1 derivatives attenuate the effects of MAG and myelin on neurite outgrowth in CGNs. (A) Monolayers of MAG-CHO and R2-CHO cells were incubated for 15 min with purified LRP1 (0.5 µM) or vehicle (veh) prior to adding CGNs. Cultures were maintained for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, **P<0.01). (B) Surfaces coated with laminin or laminin+myelin were treated for 15 min with purified LRP1 (0.5 µM) or vehicle. CGNs were then cultured for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, **P<0.01). (C) Culture wells coated with myelin and control wells were pre-treated for 15 min with shed LRP1 (0.5 µM) or vehicle. CGNs were cultured for 48 h. Neurite outgrowth was determined. The scatter plots shows neurite outgrowth in individual cells (*P<0.05). (D) Monolayers of MAG-CHO and R2-CHO cells were incubated for 15 min with 1 µM CII-Fc, CIV-Fc or Fc prior to adding CGNs for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, **P<0.01). (E) Surfaces coated with laminin or laminin+myelin were pre-treated with 1 µM purified CII-Fc, CIV-Fc or Fc for 15 min. CGNs were then added for 48 h. Neurite outgrowth was determined (means±s.e.m., n = 3, *P<0.05, **P<0.01). Scale bars: 100 µm.

Article Snippet: For expression of recombinant proteins, transfected CHO-K1 cells were cultured in Power-CHO CD medium (Lonza, Anaheim, CA).

Techniques: Incubation, Purification, Cell Culture