Figure 2

(A) Northern blot analysis for the four sRNAs (sYJ5, sYJ20 (SroA), sYJ75 and sYJ118) that were upregulated in the presence of tigecycline, and (B) bar chart illustration of the overexpressed sRNAs and (C) chromosomal locations and the directions of transcription of sYJ5, sYJ20, sYJ75 and sYJ118. A) Northern blot analysis for sYJ5, 20, 75 and 118. Image on top: all lanes marked by - were loaded with SL1344 total RNA extracted from cells grown under normal conditions (RDM, shaking, 37°C); all lanes marked by + were loaded with SL1344 total RNA extracted from cells challenged with half the MIC of tigecycline (0.125 μg/ml). Image below: representative image of the internal reference of 5S RNA levels in the same RNA samples. B) Densitometric analysis of the data from northern blot experiments of challenged / unchallenged cells with half the MIC of tigecycline. After normalisation to the 5S RNA levels, relative fold increases for sYJ5, 20, 75 and 118 were found to be 8, 2, 2, and 8 fold, respectively compared to unchallenged cells. Error bars are generated based on three independent experiments. C) The three coding sequences of sYJ5 are located in (1) SL1344_rRNA0001-rRNA0002, (2) SL1344_rRNA0014-rRNA0015 and (3) SL1344_rRNA0017-rRNA0018. The two identical copies of sYJ118 are encoded in (1) SL1344_rRNA0010-rRNA0009 and (2) SL1344_rRNA0011-rRNA0012, and the other five paralogs are found in (1) SL1344_rRNA0001-rRNA0002, (2) SL1344_rRNA0006-rRNA0005, (3) SL1344_rRNA0014-rRNA0015, (4) SL1344_rRNA0017-rRNA0018 and (5) SL1344_rRNA0020-rRNA0021.