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CORRECTIONS

CORRECTION

Abdulrazzak N., Pollet B., Ehlting J., Larsen K., Asnaghi C., Ronseau S., Proux C., Erhardt M., Seltzer V., Renou J.-P., Ullman P., Pauly M., Lapierre C., and Werck-Reichhart D. A coumaroyl-ester-3-hydroxylase Insertion Mutant Reveals the Existence of Nonredundant meta-Hydroxylation Pathways and Essential Roles for Phenolic Precursors in Cell Expansion and Plant Growth.

The authors regret that this article contains a description of immunofluorescence/confocal microscopy methodology that was not actually used for this work. In addition, this methodology was given without proper credit to Sugimoto et al. (K. Sugimoto, R.E. Williamson, G.O. Wasteneys [2000] Plant Physiol 124: 1493–1506), who developed the protocol. The correct immunofluorescence/confocal microscopy methodology used for this article is described below. The authors apologize for this error and any inconvenience it may have caused.

Immunofluorescence Visualization of the Microtubules

Roots of 2-week-old seedlings were fixed in 2% (v/v) paraformaldehyde and 0.5% (v/v) glutaraldehyde in PEMT buffer (100 mM PIPES, 4 mM EGTA, 4 mM MgSO₄, 0.05% [v/v] Triton X-100, pH 7.2) for 40 min, and rinsed in PEMT buffer three times for 10 min. Roots were postfixed in cold methanol (–20°C) for 10 min on ice, rehydrated for 10 min in 1x PBS (136 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 2 mM KH₂PO₄, pH 7.4), and treated for 20 min with NaBH₄ (1 mg/mL) diluted in 1x PBS. Fixed roots were digested for 10 min with 0.2% (w/v) pectolyase, 1% (w/v) macerozyme, 3% (w/v) caylase diluted 10 times in digestion buffer (25 mM MES, 8 mM CaCl₂, 600 mM mannitol, pH 5.5). After three washes in PBSG buffer (1x PBS, 50 mM Gly), roots were incubated for 20 min in 5% normal goat serum diluted in PBSG to saturate nonspecific sites, then incubated with primary antibodies directed against -tubulin (Molecular Probes) in PBSG at 4°C overnight and washed three times for 5 min in PBSG buffer. Samples were incubated for 1 h at room temperature with secondary antibodies coupled to Alexa Fluor 488 (Molecular Probes) and washed three times in PBSG buffer. Roots were mounted in Mowiol containing DABCO (100 mg/mL).

Observations were done using a Zeiss LSM510 confocal laser scanning microscope equipped with argon and helium/neon lasers and with a C-APOCHROMAT (x63, 1.2 numerical aperture water immersion lens). Excitation/emission wavelengths were 488/bandpass 505 to 550 nm for Alexa 488. Image processing was done using LSM510 version 2.8 (Zeiss), ImageJ (W.S. Rasband; National Institutes of Health), and Photoshop 6.0 (Adobe Systems).

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via Web of Science (3) Citing Articles via Google Scholar Google Scholar Search for Related Content