Journal: Molecular Vision

Article Title: NGF promotes cell cycle progression by regulating D-type cyclins via PI3K/Akt and MAPK/Erk activation in human corneal epithelial cells

doi:

Figure Lengend Snippet: Effect of nerve growth factor on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) and mitogen activated protein kinase (MAPK)/extracellular signal-regulated kinase (Erk) signaling pathways in human corneal epithelial cells. A : Human corneal epithelial cells (HCECs) were incubated in defined keratinocyte serum-free medium (K-SFM) without growth factors for 24 h before nerve growth factor (NGF) stimulation and then treated with NGF at 25 ng/ml for different periods. B : HCECs were treated with different concentrations of NGF for 60 min. A total of 50 μg cell lysates were analyzed for expression of the indicated genes by immunoblotting analysis. Tyr674/675 phospho-TrkA indicates the NGF signaling activity. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. For both A and B , results are representative of three independent experiments. Experiments were performed in triplicate.

Article Snippet: Antibodies against Tyr674/675 phospho-TrkA, total TrkA, Ser473 phospho-Akt, total Akt, Thr202/Tyr204 phospho-p44/42 MAPK (Erk1/2), total p44/42 MAPK (Erk1/2), cyclin D2, and anti-Rabbit IgG HRP-conjugated Secondary Antibody were from Cell Signaling Technology (Beverly, MA).

Techniques: Incubation, Expressing, Western Blot, Activity Assay

Journal: Molecular neurobiology

Article Title: Maternal Subclinical Hypothyroidism in Rats Impairs Spatial Learning and Memory in Offspring by Disrupting Balance of the TrkA/p75 NTR Signal Pathway.

doi: 10.1007/s12035-021-02403-z

Figure Lengend Snippet: Fig. 8 Effects of SCH and OHT on NGF-mediated TrkA and p75NTR signaling pathway. NGF induces TrkA signaling pathway to increase neuronal prolifera- tion. Pro-NGF, the precursor protein of NGF, can combine with p75NTR preferentially to enhance neuronal apopto- sis. CREB, cAMP response element-binding protein; JNK, C-Jun N-terminal kinase; NGF, nerve growth factor; OHT, overt hypothyroidism; p75NTR, p75 neurotrophin receptor; SCH, subclinical hypothyroidism; TrkA, tropomyosin-related kinase A

Article Snippet: The membranes were incubated overnight at 4 °C with the following antibodies: rabbit anti-NGF (1:1000 dilution; Abcam, ab52918, Cambridgeshire, UK); rabbit anti-proNGF (1:1000 dilution; Sigma-Aldrich, P5498); rabbit anti-total TrkA (1:1000 dilution; Cell Signaling Technology 2505S, USA); rabbit anti-p-TrkA (1:1000 dilution; Abcam, ab1445); rabbit anti-total ERK (1:1000 dilution; CST4695S); rabbit anti-pERK (1:2000 dilution; CST4370S); rabbit anti-total CREB (1:1000 dilution; CST9197S); rabbit anti-p-CREB (1:1000 dilution; CST9198S); rabbit anti-p75NTR (1:1000 dilution; CST8238S); rabbit anti-p53 (1:1000 dilution; CST32532S); rabbit anti-total JNK (1:1000 dilution; CST9252S); rabbit anti-p-JNK (1:1000 dilution; CST4668S); rabbit anti-Bax (1:1000 dilution; CST2772S); and rabbit anti-cleaved caspase-3 (1:1000 dilution; CST9664S).

Techniques: Binding Assay