Fig. 2

Verification of candidate methylated transcripts by meRIP. a Graphical depiction of the meRIP approach. RNA was extracted from cells, chemically fragmented, incubated with an anti-5-methylcytosine antibody or IgG, and antigen-antibody complexes were captured with protein A beads. Specific candidate RNAs (blue bars in b) were analyzed by qPCR of immunoprecipitated material, and enrichment relative to the IgG control (black bar in b) was calculated. b MeRIP shows significant enrichment of 13 out of 16 candidate transcripts. The Tbp transcript (white bar) served as a negative control, since it was not detected in our m5C dataset. Data are shown as mean ± standard error of the mean (SEM) of three independent experiments. Statistical significance was determined by unpaired t test, significance threshold p < 0.05 (*)