Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 1 | Establishment of NGF-modified NSCs and quality evaluation. (A) The design of lentiviral vectors expressing NGF (pLVX-IRES-ZsGreen-rat NGF). (B) NGF-modified NSCs highly expressed nestin and they had no immunoactivity with GFAP, CNPase, and β-tubulin3. (C) RT-PCR showed the mRNA level of NGF was significantly enhanced in NGF-NSCs. (D) Western blotting further confirmed the elevated protein concentration of NGF in NGF-NSCs. (E) Representative immunocytochemical images demonstrated high levels of NGF in NGF-NSCs neurospheres. (F) NGF-NSCs could differentiate into β-tubulin3-positive neurons, GFAP-positive astrocytes, and CNPase-positive oligodendrocytes. Scale bar = 20 µm. **p < 0.01 and ***p < 0.001.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Protein Concentration

Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 3 | Survival and differentiation of NGF-modified NSCs after transplantation and high levels of NGF at the lesion core. (A) Four weeks after NGF-NSCs transplantation, we observed that NGF + /ZsGreen + cells aggregated in the epicenter of the injured spinal cord, and several of them migrated into the peri-lesion area. In comparison, the sham and control rats did not show a positive signal of ZsGreen, and their immunoactivity of NGF was much weaker. (B) The transplanted NGF-NSCs marked by ZsGreen expressed β-tubulins3 and CNPase, which indicated the NGF-NSCs could differentiate into neurons and oligodendrocytes. (C,D) The western blotting analysis demonstrated that NGF’s protein level was much higher in the NGF-NSCs groups at day 14, 21, 28, and 35 after injury. (E) Similarly, the mRNA level of NGF exhibited the same elevation in the three groups. Scale bar, white bar 5 µm; yellow bar 200 µm. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Transplantation Assay, Comparison, Control, Western Blot

Journal: Frontiers in cellular neuroscience

Article Title: Neural Stem Cells Overexpressing Nerve Growth Factor Improve Functional Recovery in Rats Following Spinal Cord Injury via Modulating Microenvironment and Enhancing Endogenous Neurogenesis.

doi: 10.3389/fncel.2021.773375

Figure Lengend Snippet: FIGURE 4 | NGF-modified NSCs transplantation reduced the oligodendrocyte loss, attenuated astrocytosis, and demyelination in the peri-lesion segments post SCI. (A) Five weeks after SCI, the sham rats exhibited significantly higher expression of GFAP in the landscape, anterior horn, and around the central canal in the peri-lesion segments (3 mm caudal to the lesion epicenter). The graft of NGF-NSCs dramatically attenuated the abnormal GFAP expression in the same regions. (B) NGF-NSCs treatment reversed the down-regulation of CNPase in the peri-lesion segments (3 mm caudal to the lesion epicenter). after SCI. (C) The representative electoral microscopy of the white matter in the lesion epicenter showed that the integrity of the nerve sheath and corresponding axons were preserved after NGF-NSCs transplantation. Scale bar: white 200 µm; yellow 20 µm. *p < 0.05 and **p < 0.01.

Article Snippet: Following the blocking by normal goat serum, primary antibodies of Nestin (1:100, Proteintech, Wuhan, China), CNPase (1:200, Proteintech), GFAP (1:200, Cell Signaling Technology, Shanghai, China), and β-tubulin3 (1:100, Cell Signaling Technology) were added to the samples and incubated overnight at 4◦C.

Techniques: Transplantation Assay, Expressing, Microscopy

Journal: Cerebral Cortex (New York, NY)

Article Title: Expression patterns of NKCC1 in neurons and non-neuronal cells during cortico-hippocampal development

doi: 10.1093/cercor/bhac470

Figure Lengend Snippet: List of antibodies used for IHC.

Article Snippet: Anti-CNPase, monoclonal , mouse , 1:2000 or 1:5000 , AMAb91072, Atlas Antibodies.

Techniques: Affinity Purification