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. 2012 Oct 25;41(1):487–497. doi: 10.1093/nar/gks942

Figure 2.

Figure 2.

Hfq destabilizes the JM1 and JM2 duplex while it does not affect substrates with a higher GC-content. (A) UV melting experiments were carried out with gel-purified JM1-4 and JM6 double-strands in the presence or absence of 750 nM Hfq. The folded fraction α(T) = folded(T) / (folded(T) + unfolded(T)) was calculated from the melting transitions using a baseline approach. (B) The negative first derivatives of α often make small changes in the melting curve (e.g. biphasic melting behavior) more obvious. Hfq clearly shifted the melting curves of JM1 and JM2 toward lower temperatures, while its influence on the other three substrates JM3, JM4 and JM6 was insignificant. (C) The first derivative dA260 nm/dT of UV melting profiles of the jm4h substrate in the presence and absence of Hfq. The JM4h substrate displays two main melting transitions- one for the 21 base-pair long duplex (Tm,1) and one for the attached hairpin (Tm,2). Hfq clearly shifts Tm,2 toward lower temperatures while the influence on Tm,1 is comparably small.