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. 2015 Nov 13;27(11):3099–3111. doi: 10.1105/tpc.15.00750

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© 2015 American Society of Plant Biologists. All rights reserved.

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Figure 5. — LEC1 Is Required for PIF4-Mediated Hypocotyl Elongation.

(A) Hypocotyl lengths of the wild-type, 35S:PIF4, and lec1-4 35S:PIF4 plants. Five-day-old seedlings grown in the light were used for investigation. Bar = 2 mm.

(B) Hypocotyl length statistics of the wild-type, 35S:PIF4, and lec1-4 35S:PIF4 seedlings shown in (A). Data represent mean ± sd of at least 30 seedlings. Asterisks indicate significant difference between 35S:PIF4 and lec1-4 35S:PIF4 (P < 0.05, by Student’s t test).

(C) Quantitative RT-PCR analysis showing hypocotyl elongation-related genes expression in 35S:PIF4 and lec1-4 35S:PIF4. Three-day-old seedlings grown in the light were transferred to darkness for 6 h and then harvested for RNA extraction and further analysis. Relative gene expression levels were normalized against the expression of TUB2 and those in 35S:PIF4 seedlings were designated as 100%.

(D) ChIP analysis of PIF4-HA binding to G-box containing region in IAA19 and YUC8 genes upon precipitation with anti-HA antibody. Three-day-old 35S:PIF4-HA and lec1-4 35S:PIF4-HA seedlings were transferred to darkness for an additional 2 d and harvested for ChIP assay. Data represent mean ± sd of triplicates. Asterisks indicate significant changes in ChIP enrichment in lec1-4 35S:PIF4-HA compared with 35S:PIF4-HA sample (P < 0.05, by Student’s t test).

(E) Hypocotyl lengths of pER10:LEC1-MYC and pif4 pER10:LEC1-MYC transgenic line. Five-day-old seedlings grown in the light with 10 μM estradiol or mock treatment were used for investigation. Bar = 2 mm.

(F) Hypocotyl length statistics of pER10:LEC1-MYC and pif4 pER10:LEC1-MYC shown in (E). The percentage indicates the relative hypocotyl length with estradiol treatment against that with mock treatment (designated as 100%). Data represent mean ± sd of at least 30 seedlings. Asterisks indicate significant difference of relative hypocotyl length in pif4 pER10:LEC1-MYC compared with pER10:LEC1-MYC seedlings (P < 0.05, by Student’s t test).

(G) Quantitative RT-PCR analysis showing expression of hypocotyl elongation-related genes in pER10:LEC1-MYC and pif4 pER10:LEC1-MYC. Three-day-old seedlings were treated with 10 μM estradiol and immediately transferred to darkness for 3 h and then harvested for RNA extraction and further analysis. Relative gene expression levels were normalized against the expression of TUB2 and those in pER10:LEC1-MYC seedlings were designated as 100%.

(H) ChIP analysis of LEC1-MYC binding to G-box-containing region in IAA19 and YUC8 genes upon precipitation with anti-HA antibody. Three-day-old pER10:LEC1-MYC and pif4 pER10:LEC1-MYC seedlings were treated with 10 μM estradiol and immediately transferred to darkness for additional 2 d and harvested for ChIP experiment. Data represent mean ± sd of triplicates. Asterisks indicate significant changes in ChIP enrichment in pif4 pER10:LEC1-MYC compared with pER10:LEC1-MYC sample (P < 0.05, by Student’s t test).