Figure 8.

HSP gene transcript and protein levels in the wild-type, AtHSBP-knockout, and AtHSBP-overexpression lines during HSR. Six-day-old seedlings of the wild-type (WT), AtHSBP-knockout (Athsbp-1; KO), and AtHSBP-overexpression (AtHSBP-ox2; OE) lines were treated without HS (CK) or with HS at 37°C for 1 h (H1R0) and recovered from HS for 0.5 to 4 h (H1R0.5 to H1R4). A, Transcript levels of AtHSP101, AtHSP70, and AtsHSP-CI of AtsHSP18.2 and AtsHSP17.4 were determined by real-time quantitative PCR. The fold expression was normalized relative to the wild-type level at H1R0. B, Protein accumulation of HSP101, HSP70, sHSP-CI, and RPN8 was analyzed by western blot with the indicated antibodies, and the values of signal intensity relative to the wild-type sample are indicated. Similar results were obtained from three independent replicates, and one typical result is shown. RPN8 and Rubisco large subunit stained with Amido black are shown to ensure equal loading. The values of HSP, presented in percentages, were normalized to RPN8 and relative to the wild-type sample, as shown at the bottom of the panels. Transcript and protein levels were calculated from 100 seedlings of each experiment. Data are means ± sd (n = 3). *, Significant at P < 0.05 (Student's t test, as compared with the wild-type value).