Fig. 1: MC4 is activated by herbivory and interacts with AtBAG3.
From: Cleavage of Bcl-2-associated athanogene by metacaspase determines plant antiviral immunity
a Heat map depicting the expression profile of the Arabidopsis metacaspase genes (MCs) following 3 days of whitefly infestation. Expression levels are shown as log2(TPM + 1), averaged across three biological replicates. Color scale represents average intensity values in log2(TPM + 1). b Expression levels of Arabidopsis metacaspase genes were quantified by RT-qPCR after 3 days of whitefly infestation and TYLCV infection. Values are means ± standard deviation (SD) (n = 6). Data were analyzed by two-way ANOVA followed by Tukey’s test for multiple comparisons. Different lowercase letters indicated statistically significant differences (P < 0.05). The corresponding P-values can be found in the Source Data. The experiment consisted of three biological replicates. c TYLCV infection promotes the activation of MC4. MC4-Flag and MC4C139A-Flag were transiently expressed in Mock- and TYLCV-treated N. benthamiana plants. Protein detections were performed using anti-Flag (α-Flag) and anti-CP (α-CP) antibodies. Coomassie brilliant blue (CBB) stained bands of the large subunit of Rubisco (rbcL) were used as a loading control. Three biological replicates were conducted. d Whitefly infestation promotes the processing of PROPEP1-YFP, and TYLCV infection disturbs this process. Protein detection was performed using an anti-GFP antibody. CBB-stained bands of the rbcL were used as a loading control. Three biological replicates were conducted. e The interaction between MC4 and BAG3 was assessed using a yeast two-hybrid (Y2H) system. The empty vectors pGBT9 and pGAD424 were used as negative controls. f Co-immunoprecipitation (Co-IP) analysis of MC4C139A-Flag and YFP-AtBAG3 in vivo. Total protein was extracted from N. benthamiana leaves transiently expressing 35S: GUS-Flag and 35S: MC4C139A-Flag together with 35S: YFP-AtBAG3. GFP-trap beads were used to precipitate the interaction complex. Anti-GFP (α-GFP) and anti-Flag (α-Flag) antibodies were used for detection. The experiment was repeated independently three times with similar results.
