Figure 4: TbMTase37 RNAi affects steady-state RNA levels.

(a) Total cellular RNA from wild type (W), uninduced (−) and induced (+) cells was analyzed by Northern blot with radioactive oligonucleotide probes specific for each small rRNA species and a subset of cellular tRNAs were used to detect changes in steady-state RNA levels after RNAi induction. Cytoplasmic spliced leader RNA (SL RNA) was used as a normalization control, where changes in RNA levels were calculated by normalization of wild type and induced signals to SL. Cropped panels represent the radioactive signal present at the expected RNA sizes and is also the area used in quantification. The ethidium bromide stained full-length gel (rightmost panel) shows equal RNA loading with equal A₂₆₀ units in each lane. (b) The normalized ratio of induced signal was divided by the normalized wild type signal and graphed as “Fold Change”. Each bar represents an average of 3 independent gels and blotting experiments with error bars representing standard error of mean. Statistical analysis was performed using a one-sample t-test. An asterisk (*) represents a significant (P < 0.05) difference from 1.0 (no change, dashed line).