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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).

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