Journal: Biomedicines

Article Title: Histone H3 Acetylation Is Involved in Retinoid Acid-Induced Neural Differentiation through Increasing Mitochondrial Function

doi: 10.3390/biomedicines11123251

Figure Lengend Snippet: RA promotes histone acetylation through RA signaling during neural differentiation of SH-SY5Y cells. ( a – c ) RT-qPCR analysis of relative mRNA expression of RARα , RARβ , and RARγ in SH-SY5Y cells after RA treatment for 3 and 7 days. n = 3; ** p < 0.01; one-way ANOVA. ( d ) Co-immunoprecipitation was performed in SH-SY5Y cells cultured in maintenance medium or stimulated with RA for 3 and 7 days. Immunoprecipitation was conducted using anti-RARα followed by Western blot analysis of PCAF, CBP, and P300. ( e – g ) Western blot analysis of H3K14ac and H3ac in SH-SY5Y cells treated with DMSO or RA or pretreated with BMS195614/LE135 for 24 h followed by RA for 3 days. n = 3; * p < 0.05; one-way ANOVA.

Article Snippet: To identify the roles of key enzymes and receptors during neural differentiation, the HAT activity inhibitor C646 (Sigma, St. Louis, MO, USA), RARα antagonist BMS195614, and RARβ antagonist LE135 (Glpbio, Montclair, CA, USA) were supplemented during the RA treatment.

Techniques: Quantitative RT-PCR, Expressing, Immunoprecipitation, Cell Culture, Western Blot

Journal: Biomedicines

Article Title: Histone H3 Acetylation Is Involved in Retinoid Acid-Induced Neural Differentiation through Increasing Mitochondrial Function

doi: 10.3390/biomedicines11123251

Figure Lengend Snippet: Inhibition of RA signaling downregulates mitochondrial function and blocks neural differentiation. SH-SY5Y cells were treated with DMSO or RA or pretreated with BMS195614/LE135 for 24 h followed by RA for 3 days. ( a – d ) RT-qPCR analysis of relative mRNA expression of mitochondrial function-related genes ( NDUFS3 , ANT1 , ANT2 , ANT3 ). n = 3; * p < 0.05, ** p < 0.01; one-way ANOVA. ( e ) Changes in morphology and expression of neuronal differentiation markers Tuj1 (green) and MAP2 (red). Scale bar, 50 µm. Images in the black box show cells at a higher magnification. Red or white arrows show representative changes in morphology. ( f – h ) Differentiation rates and mean fluorescence intensity of Tuj1 and MAP2. n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one-way ANOVA. ( i , j ) RT-qPCR analysis of relative mRNA expression of the neural differentiation markers Tuj1 and MAP2 . n = 3; * p < 0.05, ** p < 0.01; one-way ANOVA.

Article Snippet: To identify the roles of key enzymes and receptors during neural differentiation, the HAT activity inhibitor C646 (Sigma, St. Louis, MO, USA), RARα antagonist BMS195614, and RARβ antagonist LE135 (Glpbio, Montclair, CA, USA) were supplemented during the RA treatment.

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Fluorescence