Fig. 3

Validation of the differential expression observed by RNA-seq. The expression of randomly selected DEGs [12 DEGs identified with EdgeR tool and 8 DEGs identified with NOISeq tool, 4 of which are deregulated with both approaches (common DEGs)] was analyzed by rt-qPCR in ruminal wall of dairy buffaloes fed a TMR + green feed (Treated) in comparison with animals fed the TMR diet (Control). Data were normalized to β-actin and are reported as mean fold increases ± standard error of the mean (SEM) relative to transcript levels of control group samples. Six control and six treated animals were used, and two independent qPCR experiments were performed. * p < 0.05; * p < 001; *** p < 0.001 (one-tailed Student’s t-test). §: rt-qPCR data conflicting with RNA-seq results