Journal: Frontiers in Cell and Developmental Biology

Article Title: NLGN3 Upregulates Expression of ADAM10 to Promote the Cleavage of NLGN3 via Activating the LYN Pathway in Human Gliomas

doi: 10.3389/fcell.2021.662763

Figure Lengend Snippet: NLGN3 activated the LYN pathway to upregulate ADAM10, which in turn promoted cleavage of NLGN3. (A) The NLGN3 plasmid carrying the Flag fragment was transfected into U251 cells with an empty vector as the control. Western blot was performed to detect the Flag label. SS = signal sequence; M = maker. (B) The level of phosphorylation of AKT and LYN was detected after treatment with a NLGN3 plasmid, with an empty vector as the control. WT = wild type. (C) U251 cells were transfected with LYN or treated with bafetinib along with NLGN3 plasmid transfection, with an empty vector or DMSO as the control. Western blot was performed to detect the Flag label. WCL = whole cell lysates. (D) qPCR was performed to detect the expression of ADAM10 in U251 cells. (E) Western blot was performed to detect the expression of ADAM10 in U251 cells. ADAM10 expression was upregulated by LYN and inhibited by bafetinib. (F) Compared with the control cells, LYN overexpression increased the level of cleaved NLGN3, whereas decreased cleaved NLGN3 was observed after treatment with the ADAM10 inhibitor compared with LYN overexpression cells. (G) Overexpression of ADAM10 increased cleaved NLGN3 levels, but there was no significant change of cleaved NLGN3 levels after treatment with LYN inhibitors compared with ADAM10 overexpression group. * p < 0.05; ** p < 0.01.

Article Snippet: The ADAM10 primer (F: 5′-ATGGTGTTGCTGAGAGTGTT-3′; R: 5′-GACTGCTCTTTTGGCACGC-3′) was purchased from Ribobio Co., Ltd. (Guangzhou, China).

Techniques: Plasmid Preparation, Transfection, Western Blot, Sequencing, Expressing, Over Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: NLGN3 Upregulates Expression of ADAM10 to Promote the Cleavage of NLGN3 via Activating the LYN Pathway in Human Gliomas

doi: 10.3389/fcell.2021.662763

Figure Lengend Snippet: ADAM10 promoted the migration and invasion of glioma cells. (A) Correlation analysis between expression of ADAM10 and NLGN3 in glioma and normal brain tissue was performed using GEPIA. (B) Correlation analysis between expression of ADAM10 and LYN in glioma and normal brain tissue was performed using GEPIA. (C) The expression of ADAM10 in LGG and GBM was analyzed on GEPIA. (D) U87 cells were treated with ADAM10 inhibiter, GI254023X (10 μM), and CCK-8 assay was performed to detect the proliferation of each group of cells. (E,F) ADAM10 inhibiter, GI254023X, suppressed the migration of U87 cells. (G,H) GI254023X inhibited the invasion of U87 and U251 cells. (I) The relationship between ADAM10 and prognosis of patients with LGG and GBM was analyzed on GEPIA. * p < 0.05.

Article Snippet: The ADAM10 primer (F: 5′-ATGGTGTTGCTGAGAGTGTT-3′; R: 5′-GACTGCTCTTTTGGCACGC-3′) was purchased from Ribobio Co., Ltd. (Guangzhou, China).

Techniques: Migration, Expressing, CCK-8 Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: NLGN3 Upregulates Expression of ADAM10 to Promote the Cleavage of NLGN3 via Activating the LYN Pathway in Human Gliomas

doi: 10.3389/fcell.2021.662763

Figure Lengend Snippet: Schematic illustration of the positive feedback circle, s-NLGN3/LYN/ADAM10. Secreted NLGN3 derived from neurons and glioma cells activates LYN and then up-regulates ADAM10 expression, which can cleave NLGN3 to promote its secretion.

Article Snippet: The ADAM10 primer (F: 5′-ATGGTGTTGCTGAGAGTGTT-3′; R: 5′-GACTGCTCTTTTGGCACGC-3′) was purchased from Ribobio Co., Ltd. (Guangzhou, China).

Techniques: Derivative Assay, Expressing