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. 2023 Dec 29;11(2):uhad282. doi: 10.1093/hr/uhad282

Figure 7.

Figure 7

CsbHLH1 is localized in the nucleus of the cell and has transactivating activity, and it binds to the promoter of CsTCS1. A Subcellular location of CsbHLH1 in tobacco leaf; scale bars = 10 μm. B Transcriptional transactivation in tobacco. C Transcriptional activity of CsbHLH1 in yeast cells. D Protein purification to obtain a fusion protein of MBP-CsbHLH1. M, protein marker; 1 and 2, supernatant and precipitation of 0.5 mmol/l IPTG-induced protein; 3 and 4, supernatant and precipitation of 0.7 mmol/l IPTG-induced protein; 5, 6, and 8, purified CsbHLH1 recombinant protein washing solution. 7, concentrated CsbHLH1 recombinant protein washing solution. E EMSA. From left to right: negative control; positive control; CsTCS1 probe; CsTCS1 probe bound to CsbHLH1 protein; CsTCS1 mutant probe; CsTCS1 mutant probe and CsbHLH1 protein; CsbHLH1, CsTCS1 mutant probe, and 10× CsTCS1 competitive probe; CsTCS1 mutant probe and 50× CsTCS1 competitive probe; CsTCS1 mutant probe and 100× CsTCS1 competitive probe. The 10× and 50× CsTCS1 competitive probes can compete with part of the strip, and the 100× CsTCS1 competitive probes can compete with all the strip. F Gene expression of CsbHLH1 in leaves after As ODN_CsbHLH1 treatment expression. G Caffeine content in young shoots treated with water and As ODN_CsbHLH1. H Relative expression of CsTCS1 after inhibition of CsbHLH1.