Figure 4

Differential gene expression profiling and verification of B cells immunized with 0.05 and 0.25 mg/mL BP5. Volcano plots and venn diagram of DEG in B cells from the spleen cells of mice immunized with or without BP5. A Volcano plot of DEG in response to 0.05 mg/mL BP5. Each dot was a gene. Blue dots: down-regulated, P value < 0.05 and log₂ Fold Change ≥ 1.5, Orange dots: up-regulated, P value < 0.05 and log₂ Fold Change ≥ 1.5. Grey dots did not fit these criteria. B Volcano plot of DEG in response to 0.25 mg/mL BP5. Each dot was a gene. Blue dots: down-regulated, P value < 0.05 and log₂ Fold Change ≥ 1.5, Orange dots: up-regulated, P value < 0.05 and log₂ Fold Change ≥ 1.5. Grey dots did not fit these criteria. C Venn diagram showing the number of DEG between 0.05 and 0.25 mg/mL BP5. These genes were shared between both comparisons. D-E Fold-change expression of related genes in B cells from mice spleen immunized with BP5. qRT-PCR was performed for each gene to validate the RNA-Seq results. Seven genes were selected from the DEG results from the comparison of 0.05 and 0.25 mg/mL BP5, with FDR 0.05 and log₂ fold change cut-off. The housekeeping gene β-actin was used to normalize relative expression. Statistically significant differences were determined using the student t-test.