FIG 4.

Northern blot analysis of sRNA teg49 in S. aureus strains in various phases of growth. (A, E) Northern blot assay detection of sRNA teg49. Total RNAs (10 μg) from ΔcshA (A) and ΔsigB (E) mutants of strain Newman derivative ALC6094 grown in TSB medium were separated in a 1.5% agarose gel and then transferred to an H⁺-bond membrane for probing. sRNA teg49 was detected with a teg49 probe. (B) Primer extension analysis of sRNA teg49 in a cshA mutant of ALC6094. Primer extension assays were performed with RNAs purified from the parental (ALC6094), ΔcshA mutant, and complemented strains grown to exponential phase (OD₆₅₀ of ∼0.7) in TSB medium with oligonucleotide PA-13 (Table 2). ctrl, control; wt, wild type. (C, D) Quantification of sarA P3 and teg49 transcripts in the cshA mutant, parental, and complemented strains from panel B. Transcripts sarA P3 (C) and sRNA teg49 (D) were quantified with ImageJ (NIH). These experiments were repeated at least three times, and one set of typical experiments is shown.