ATG16 | Autophagy-related protein 16

Detection of lace plant Atg16 in lace plant leaves.RNA and protein were extracted and probed for Atg16 from imperforate (I), pre-perforation (P), window (W), or mature (M) lace plant leaf stages (A-D) or only window and mature leaves from plants treated with either 5 μM rapamycin (Rap), 1 μM wortmannin (Wrt), and 1 μM concanamycin A (ConA) compared to DMSO control (E-H). (A and E) The mean levels of Atg16 mRNA in different lace plant leaf stages were determined by qRT-PCR and normalized to lace plant α-tubulin levels. Protein extracts and a molecular protein standard (L) were resolved by SDS–polyacrylamide gels and blotted to nitrocellulose membranes. (B and F) Membranes and protein lanes were stained with Ponceau-S to serve as loading control before subsequent detection of the presence or absence of ~56 kDa sized Atg16 protein bands with anti-Atg16 antibody (C and G). (D and H) Immunoreactive protein bands were quantitated, and the ratio of Atg16 band intensity to the Ponceau lane signal was averaged. The experiments were performed in triplicate. Means not sharing any letter are significantly different. One-way ANOVA (A, D and H), or Two-way ANOVA, Tukey test (E, P < 0.05; n = 3). Error bars represent the SE.

Detection of lace plant Atg16 in lace plant leaves.RNA and protein were extracted and probed for Atg16 from imperforate (I), pre-perforation (P), window (W), or mature (M) lace plant leaf stages (A-D) or only window and mature leaves from plants treated with either 5 μM rapamycin (Rap), 1 μM wortmannin (Wrt), and 1 μM concanamycin A (ConA) compared to DMSO control (E-H). (A and E) The mean levels of Atg16 mRNA in different lace plant leaf stages were determined by qRT-PCR and normalized to lace plant α-tubulin levels. Protein extracts and a molecular protein standard (L) were resolved by SDS–polyacrylamide gels and blotted to nitrocellulose membranes. (B and F) Membranes and protein lanes were stained with Ponceau-S to serve as loading control before subsequent detection of the presence or absence of ~56 kDa sized Atg16 protein bands with anti-Atg16 antibody (C and G). (D and H) Immunoreactive protein bands were quantitated, and the ratio of Atg16 band intensity to the Ponceau lane signal was averaged. The experiments were performed in triplicate. Means not sharing any letter are significantly different. One-way ANOVA (A, D and H), or Two-way ANOVA, Tukey test (E, P < 0.05; n = 3). Error bars represent the SE.