Journal: PLoS ONE

Article Title: Neuroinductive properties of mGDNF depend on the producer, E . Coli or human cells

doi: 10.1371/journal.pone.0258289

Figure Lengend Snippet: All dissociated embryonic ganglia were immunohistochemically stained for β3-tubulin to confirm the neuronal origin of the processes; and all processes were β3-tubulin-positive. ( a ) control, no GDNF; ( b ) mGDNF/Coli; ( c) Pro-mGDNF/Coli. Scale, 500 μm.

Article Snippet: After 10 days of culture, spinal ganglia were fixed in 4% formaldehyde in PBS at 4°C for 20 min and washed with PBS for 5 min. For immunohistochemical analysis, the primary rabbit polyclonal antibodies against β3-tubulin (Abcam, UK) were diluted 1:100 with PBS containing 0.3% Triton X-100 (Sigma, USA), and 2% normal donkey serum.

Techniques: Staining, Control

Journal: PLoS ONE

Article Title: Neuroinductive properties of mGDNF depend on the producer, E . Coli or human cells

doi: 10.1371/journal.pone.0258289

Figure Lengend Snippet: (a) Preliminary evaluation of MPP+ toxic effect on human neuroblastoma SH‐SY5Y cell survival. The dose-dependence and high reproducibility were demonstrated. (b) Analysis of β3-tubulin expression in cultured SH-SY5Y neuroblastoma cells exposed to the factors: 1, recGDNF; 2, Pro-mGDNF/Coli; 3, mGDNF/Coli. (c) and (d) Effect of the obtained recombinant GDNF factors on SH-SY5Y neuroblastoma cells after the exposure to МРР+ for 30 h. SH-SY5Y was incubated with different concentrations of GDNF factors (10, 20, 50 ng/mL), (c) 2 mM МРР+; (d) 3 mM МРР+. (e) Effect of recGDNF, mGDNF/Coli, and Pro-mGDNF/Coli on the viability of SH-SY5Y neuroblastoma cells at 48-h exposure to MPP+; gdnf-, recombinant factor recGDNF derived from the full-length GDNF; m, mGDNF/Coli; pro, Pro-mGDNF/Coli. Data represents the mean ± SEM (for (a), (b) and (e) *p<0,05 vs. control) (N = 3).

Article Snippet: After 10 days of culture, spinal ganglia were fixed in 4% formaldehyde in PBS at 4°C for 20 min and washed with PBS for 5 min. For immunohistochemical analysis, the primary rabbit polyclonal antibodies against β3-tubulin (Abcam, UK) were diluted 1:100 with PBS containing 0.3% Triton X-100 (Sigma, USA), and 2% normal donkey serum.

Techniques: Expressing, Cell Culture, Recombinant, Incubation, Derivative Assay, Control