Fig 2: The expression of heat stress response transcripts is altered in the absence of TFIIS. Relative expressions of (A) HsfA2, (B) HSP101, (C) HSP70, (D) HSP90-1, (E) sHSP18.1 and (F) sHSP22.0 transcripts as measured by qRT-PCR analysis; treatment conditions as in Figure 4; bars represent standard errors based on at least three biological replicates; P-values based on two-tailed Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001).

Fig 3: Time-course of HSP101 gene expression and HSP101 protein accumulation after treatment with chemical inducers or moderate heat. (A) Seedlings were treated with the HSP90-inhibitors celastrol (CEL, 20 µM), radicicol (RAD, 50 µM), geldanamycin (GDA, 50 µM), and the RES cyclopentenones (PGA1 and OPDA, 75 µM) for the times indicated. HSP101 mRNA levels were determined by qPCR and normalized to AtSAND; data represent means ±SD, n=3. (B) Relative HSP101 protein levels were evaluated by dot blot analysis and shown as a percentage of seedlings subjected to 37 °C heat treatment for 4 h. Data represent means of three technical replicates of a representative experiment ±SD.

Fig 4: Heat- and wound-induced jasmonate accumulation and HSP101 expression. Time-course of jasmonate accumulation in seedlings shifted to 37 °C (A, C) and wounded wild-type and dde2 leaves from 6-week-old plants (B, D). Relative HSP101 expression (as determined by qPCR and normalized to AtSAND expression) in wild-type and mutant seedlings after 1 h at the temperatures indicated (E) or after wounding (F). Data represent means ±SD, n=3.

Fig 5: Differential transcription of heat-induced genes in the wild type (WT) and transgenic lines (OE1 and OE2). 10-day-old seedlings were incubated at a specified temperature for 2 h and then used to extract RNA. Transcription levels of thermotolerance-related genes HsfA2 (AT2G26150), HsfB1 (AT4G36900), Hsp60 (AT2G28000), Hsp70 (AT3G09440), Hsp101 (AT1G74310), sHsp class I (AT3G46230) and sHSP class II (AT5G12030). Quantitative analysis was performed by qRT-PCR. Actin2 was used as the reference gene.