Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Loss of NgBR causes neuronal damage through decreasing KAT7-mediated RFX1 acetylation and FGF1 expression

doi: 10.1007/s00018-025-05660-6

Figure Lengend Snippet: NgBR deficiency leads to the downregulation of FGF1 expression and secretion. A Expression clustering heatmap analysis of differentially expressed genes related to the PI3K-AKT pathway shown in Fig. B. B‒D SH-SY5Y cells were transfected with the indicated siRNAs for 48 h. Then, the samples were subjected to qRT‒PCR ( B ) or immunoblotting ( C ). The relative protein level of FGF1 to that of β-actin in ( C ) was analyzed as shown in D. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001. E SH-SY5Y cells were transfected with Flag or Flag-NgBR for 48 h, after which the cell lysates were subjected to immunoblot analysis. F The protein levels of FGF1 relative to those of β-actin in ( E ) were analyzed. n = 3, ** P < 0.01. G N2a cells were transfected with the indicated siRNAs for 48 h. Then, the cell lysates were subjected to immunoblot analysis. H The protein level of FGF1 relative to that of β-actin in ( G ) was analyzed. n = 3, * P < 0.05, ** P < 0.01. I MES23.5 cells were transfected with the indicated siRNAs for 48 h. Then, the cell lysates were subjected to immunoblot analysis. J The protein level of FGF1 relative to that of β-actin in ( I ) was analyzed. n = 3, * P < 0.05. K Nus1 flox/flox primary cortical neurons were infected with LV-Ctrl or LV-Cre and cultured for 6 days. Then, the cell lysates were subjected to immunoblot analysis. L The protein level of FGF1 relative to that of β-actin in ( K ) was analyzed. n = 3, ** P < 0.01. M-N SH-SY5Y cells were transfected with the indicated siRNAs for 24 h. Then, the cells were cultured with serum-free medium for 24 h, and the FGF1 content in the culture medium ( M ) and cell lysate ( N ) was detected via an ELISA kit. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: SH-SY5Y cells were inoculated into 24-well plates and treated with siRNAs against NUS1 for 24 h. Then, the cells were cultured with serum-free medium for 24 h, and 100 μL of the FGF1 content in the culture medium or in cell lysate was measured with an ELISA kit (EK0339, BOSTER, Wuhan, China).

Techniques: Expressing, Transfection, Western Blot, Infection, Cell Culture, Enzyme-linked Immunosorbent Assay