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Figure 4 | BMC Plant Biology

Figure 4

From: Histone acetylation associated up-regulation of the cell wall related genes is involved in salt stress induced maize root swelling

Figure 4

High-salinity increased histone acetylation levels. (A, B) The histones H3K9 and H4K5 acetylation levels were increased after 200 mM NaCl treatment. Nuclei from six-day-old seedlings untreated (Control) or treated with 200 mM NaCl for 24, 48, 72 and 96 h were subjected to immunofluorescence using antibodies against H3K9Ac (A) and H4K5Ac (B). The ‘DAPI’ panel displays DAPI-stained DNA images pseudo-colored with blue, the ‘H3K9Ac’ and ‘H4K5Ac’ panels show immunostained images pseudo-colored with green, and the ‘Merge’ panel indicats a combination of blue and green signals. Bar = 10 μm. (C, D) Histogram displaying the mean gray values of the immunostaining signals for H3K9Ac (C) and H4K5Ac (D). The mean gray values for H3K9Ac and H4K5Ac were increased by approximately 40% to 60% after treatment with 200 mM NaCl. More than 300 nuclei were analyzed and error bars represent the standard error of the mean gray value. (E) High-salinity affects H3K9Ac and H4K5Ac protein levels. The antibodies specific for H3K9Ac and H4K5Ac were employed to analyze histones extracts from seedlings untreated (Control) and treated with 200 mM NaCl for 24, 48, 72 and 96 h. Histone H3 was applied as an equal loading control. The average values of abundance index of H3K9Ac and H4K5Ac were indicated in each lane (Mean ± SD). After treatment with 200 mM NaCl, an increase in global H3K9Ac and H4K5Ac were observed at the indicated times. (F, G) Expression patterns of ZmHATB (F) and ZmGCN5 (G) genes at different time points were determined by RT-PCR in maize seedlings under salt stress conditions. The y-axis indicates relative expression value and the x-axis indicates the hours after salt stress. The relative quantity was calculated using the beta actin gene as an internal reference. The relative expression value of 0 h was assigned as 1. Each experiment was repeated three times and relative expression values are shown as the average values ± SD (the standard deviation). White bars, no treated seedlings; Black bars, seedlings treated with 200 mM NaCl.

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