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. 2022 Oct 20;13:943627. doi: 10.3389/fphar.2022.943627

FIGURE 4.

FIGURE 4

Longer term B-27 exposure leads to SH-SY5Y cell cycle arrest. SH-SY5Y cells were cultured in DMEM/F-12 only, or DMEM/F-12 supplemented with either 10% FBS, 2% B-27 or 5% B-27 for (A) 48 h or (B) 96 h. qRT-PCR was performed to measure mRNA expression of cell cycle markers CDK6 and CDK1, and apoptosis marker CASP3. Relative expression of CDK1, CDK6, and CASP3 (standardized to GAPDH and POLR2A) normalized to the DMEM/F-12 + 10% FBS culture conditions. Data presented as mean ± SEM. N = 3 independent biological samples with two to three technical replicates each. Statistical significance against the DMEM/F-12 + 10% FBS condition was determined using ordinary one-way ANOVA and Tukey’s multiple comparison tests in GraphPad Prism version 9.0.0 for Mac, GraphPad Software, San Diego, California, United States, www.graphpad.com. ns = not significant; *p < 0.05; **p < 0.01; ****p < 1 × 10–4. CASP3, Caspase 3; CDK1, Cyclin dependent kinase 1; CDK6, Cyclin dependent kinase 6; DMEM/F-12, Dulbecco’s modified eagle medium/nutrient mixture F-12 with GlutaMAX supplement; FBS, fetal bovine serum; GAPDH, Glyceraldehyde-3-phosphate dehydrogenase; mRNA, messenger RNA; POLR2A, RNA polymerase II subunit A; SEM, standard error of the mean.