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. 2014 Feb 21;42(8):5270–5279. doi: 10.1093/nar/gku157

Figure 3.

Figure 3.

Central pairing is required for miR398-mediated repression of BCBP. (A) Pairing of miR398a with the corresponding complementary sites of both BCBP and an miR398-resistant form of BCBP (2mBCBP) containing two nucleotides (in blue) that disrupt the complementarity. (B) qRT-PCR quantification of BCBP mRNAs in transgenic plants (T3 generation) carrying either a BCBP-unmodified cDNA or a 2mBCBP version, driven by its own promoter (proBCBP). Plants were grown for 12 days in vitro, with (CuSO4 0.3 µM) or without copper added to the medium. MIR398b/c genes are transcribed in the absence of copper. Total RNAs were extracted in bulk (10–15 plants) to prepare the corresponding cDNAs and perform the qRT-PCR. Results are presented for five independent transgenic lines carrying proBCBP:BCBP (lines #7, #13, #17, #23 and #27) and five independent lines for proBCBP:2mBCBP (lines #5, #6, #13, #16 and #19). Error bars indicate SD (n = 5). One microgram of total RNAs was analyzed by northern blot to control miR398 levels.