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Fig. 2 | BMC Plant Biology

Fig. 2

From: Clarification of the dispensability of PDX1.2 for Arabidopsis viability using CRISPR/Cas9

Fig. 2

The PDX1.2 protein accumulates upon heat stress. a Confocal micrographs (z slices) of cotyledons and roots of 8-days-old Arabidopsis expressing the PDX1.2-YFP fusion protein under the control of the upstream region of PDX1.2 (pPDX1.2::PDX1.2-YFP), in the absence (−HS) and presence of heat stress (+HS). L1 and L3 refer to independent lines. Heat stress was induced by incubating seedlings for 3 h at 37 °C. YFP expressed alone under the control of the CaMV 35S promoter (35S::YFP) and non-transgenic wild type (Col-0) are also shown for comparison. Scale bars: 20 μm. A color scale bar of fluorescence intensity is shown on the right. b Fluorescence intensities (arbitrary units) measured in cotyledons and in roots. Note that 35S-YFP plants were imaged with different acquisition parameters than the other lines, making the absolute values measured in this line not comparable with those measured in the others. The data are the average of 8–67 tissues from at least 2 plants per genotype, tissue and condition (see methods) and are represented as the average ± SE. Statistical differences were calculated by a two-tailed Student’s t test for genotype/tissue with and without heat stress and are indicated by an asterisk for P < 0.05. c Immunochemical analysis of 8-days-old whole seedlings of independent pPDX1.2::PDX1.2-YFP lines (L1 and L3) compared to wild type (Col-0) using an antibody against GFP (α-GFP). An antibody against actin (α-Actin) was used as a loading control. The arrows point to labelled bands at 62 kDa and 42 kDa, the expected sizes of the PDX1.2-YFP fusion protein and actin, respectively. Samples correspond to treatment with heat stress (+HS) or non-treatment (−HS) as shown for part (a)

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