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Neuro2a cells co-transfected with FLAG-SOD1 and HA-CHGB were immunostained using anti-HA and anti-FLAG (C) or anti-misf SOD1 antibodies (B8H10) (D).
However, the expression of CHGB (WT, L413 and R230) and mouse Cg B did not promote the secretion of endogenous mouse SOD1 in cells (data not shown).
Since previous studies demonstrated a role for CHGB in supporting neurite outgrowth of cultured cells (34–36), we examined the effects of CHGB transgenic mouse lines that overexpressed human CHGB m RNA species at similar levels in the spinal cord as assessed by quantitative real time RT-PCR.
However, these results have not been corroborated by other groups that reported a lack of association of , detected only in few ALS cases but not in control individuals (28).
We carried out transient co-expression assays in Neuro2a cells using plasmid vectors coding for mouse chromogranin B (m Cg B) or various human CHGB species tagged with hemagglutinin (HA) at the carboxy terminus together with vectors coding for wild type (WT) or SOD1.
Neuro2A cells were co-transfected with FLAG-tagged SOD1 and HA-tagged CHGB.