-Tubulin Is Essential for Acentrosomal Microtubule Nucleation and Coordination of Late Mitotic Events in Arabidopsis

Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - The strength of the Phenotype and the Strength of GUS Staining of Transformants Expressing the Dexamethasone Inducible γ-Tubulin RNAi Construct (A) GUS staining of roots with weak phenotype, (B) mild phenotype (C) strong phenotype, sometimes the correlation between the strength of phenotype and intensity of GUS expression is clearly visible on one plant, arrow marks aberrant root of strong phenotype. Bar = 1 mm (D) GUS activity extracted from roots with the weak phenotype (A), mild phenotype (B), and strong phenotype (C). For each phenotype - weak (A), mild (B) and strong (C) four samples were analyzed. Error bars represent SD.
• Supplemental Figure 2 - The lack of MT Polymerization from Extracts Immunodepleted for γ-Tubulin Observed on Cover Slip in in vitro Polymerization Assays On slide in vitro MT polymerization assays from control, mock depleted and γ-tubulin immunodepleted extracts. The same extracts were analyzed for γ-tubulin levels by protein gel blot (data shown in Figure 2C). MTs polymerized on slides were observed directly with DIC optics or after visualization by immunofluorescence labeling for α-tubulin, with anti-mouse Cy3 conjugated secondary antibody. Bars = 10 μm.
• Supplemental Figure 3 - MTs in Root Hairs of the Control and RNAi Plants α-tubulin labeling (green) and DAPI labeling (blue). Compared to the relatively dense MTs in the control root hairs (A), short bundled MTs are observed in the vicinity of the nucleus in root hairs 5 days after γ-tubulin RNA induction.