To study the effect of hormones on REEs accumulation, one set of asynchronous cultures of G. sulphuraria was cultivated with CFL and hormones for 24 h. For this study two synthetic plant hormones, 6-Benzylaminopurine (BAP-Cytokinin family) and 1-Naphthaleneacetic acid (NAA-Auxin family) (Sigma-Aldrich) were used at a final concentration of 5 mg L−1 for this study. At the end of the experiment, cultures with and without plant hormones were harvested by centrifugation (3,000 rpm, 5 min), freeze dried and analyzed by ICP-MS.
1 naphthaleneacetic acid
Manufactured by Merck Group
Sourced in Germany, Poland, United States
1-Naphthaleneacetic acid is a chemical compound that is commonly used in laboratory settings. It is a white crystalline solid with the molecular formula C₁₁H₉O₂. The primary function of 1-Naphthaleneacetic acid is to serve as a plant growth regulator, helping to control various physiological processes in plants.
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Lab products found in correlation
6 benzylaminopurine, by Merck Group (3 mentions)
Kinetin, by Merck Group (2 mentions)
1 na pp1, by Euromedex (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
2 4 dichlorophenoxyacetic acid, by Merck Group (2 mentions)
Acetone, by Avantor (1 mentions)
Indole 3 acetic acid, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Avantor (1 mentions)
Murashige and skoog, by Merck Group (1 mentions)
Mercury 2 chloride hgcl2, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Gibberellic acid, by Merck Group (1 mentions)
Ck 6 benzylaminopurine ba, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Whwlqlkpgqpmy, by Merck Group (1 mentions)
1 10 phenanthroline monohydrate, by Merck Group (1 mentions)
Actinomycin d, by Merck Group (1 mentions)
N0640, by Merck Group (1 mentions)
3 indoleacetic acid, by Merck Group (1 mentions)
Diphenyleneiodonium dpi, by Merck Group (1 mentions)
16 protocols using 1 naphthaleneacetic acid
1
Hormonal Influence on REEs Accumulation in G. sulphuraria
2
Plant Growth Regulation: A Comprehensive Protocol
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6-Benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), kinetin (KIN), Murashige and Skoog (MS) basal medium, and mercury (II) chloride (HgCl2), were acquired from Sigma-Aldrich, Germany. Dimethyl sulfoxide, acetone, and additional solvents and reagents were bought from VWR International, Belgium.
3
Maize mTERF Expression Regulation
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To verify the expression regulation of mTERF genes in maize under hormone and salt [sodium chloride (NaCl, 200 mM) and aluminum chloride (AlCl3, 200 mM)] treatments, the two-leaf B73 seedlings were cultured in water under the following chemical treatments, ABA (100 μM) and 1-Naphthaleneacetic acid (NAA, a synthetic plant hormone in the auxin family) (100 μM) (Sigma-Aldrich, Shanghai), respectively. Samples were collected at 0.5, 1, 2 and 4 h after the above treatment, with three biological replicates per sample. To investigate effects of light on expression of maize mTERFs, etiolated B73 seedlings cultured in darkness were placed under continuous light, while normal-cultured B73 seedlings cultured under 16 h light/8 h dark were placed in darkness after 24 h of continuous light. Samples were collected at 1, 2, 4 and 8 h after light or dark treatment, with three biological replicates per sample. Total RNAs of collected samples were isolated with Trizol reagent (Invitrogen, USA) according to the manufacturer’s instructions and quantified by NanoDrop 2000 spectrophotometer (ThermoFisher, USA) before cDNA synthesis.
4
Exogenous Hormone Application in Plants
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All exogenous applications of the synthetic CK 6-benzylaminopurine (BA) (Sigma-Aldrich, St Louis, MO, USA) were performed by spraying or immersing the plants for 5 min. Absicic acid (ABA, 100 µM), 1-Naphthaleneacetic acid (NAA, 100 µM), gibberellic acid (GA, 100 µM) paclobutrazol (paclo, 10 mg/mL), all from Sigma-Aldrich, and Ethylene (Ethrel, Bayer Cropscience) were applied by spraying. All hormone treatments included the surfactant Tween 20 (100 μl l−1).
5
Auxin Regulation of Root Development
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The plant seeds from the different genotypes were sown onto solid growth medium, and vertically grown as described above. Plates were scanned using a flatbed scanner every day from the fourth to the 10th dag. The roots were analysed at different developmental stages through image analysis (Fiji – ImageJ bundle software). The experiments were performed at least three times and each sample comprised 25 seedlings.
For N-1-naphtylphtalamic acid (NPA) and auxin treatments, 5-d-old seedlings were transferred to growth medium supplemented with different concentrations of NPA, IAA, or 1-Naphthaleneacetic acid (NAA; Sigma-Aldrich). After 5 d of treatment, the primary root length was measured as described above.
For N-1-naphtylphtalamic acid (NPA) and auxin treatments, 5-d-old seedlings were transferred to growth medium supplemented with different concentrations of NPA, IAA, or 1-Naphthaleneacetic acid (NAA; Sigma-Aldrich). After 5 d of treatment, the primary root length was measured as described above.
6
Imaging Meiosis with Aurora and Condensin Inhibition
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The ark1-as3 analog-sensitive allele (Koch et al., 2011 (link)) was used to perform Ark1 inhibition. To inhibit Aurora in meiosis I or II, the imaging media was supplemented with 10 µM 1-NA-PP1 (Euromedex). However, to specifically inhibit Aurora kinase in meiosis II, image acquisition started exactly 10 min after the spreading of cells on the imaging media containing 1-NA-PP1, and only cells starting meiosis II were filmed. This protocol allows imaging of cells that underwent normal chromosome segregation in meiosis I.
Condensin depletion was performed using the cut14-so strain previously described (Kakui et al., 2017 (link)) that combines transcriptional repression of cut14 under the nmt81 thiamine-repressible promoter and Cut14 depletion using an Auxin-Inducible Degron (AID) system. To induce condensin depletion, the imaging media was supplemented with 0.5 mM of 1-Naphthaleneacetic acid (Sigma) and 5 μg/ml of thiamine. For condensin depletion in meiosis I, image acquisition was started 3 h after spreading the cells on the imaging media containing Naphthaleneacetic acid. For condensin depletion in meiosis II, image acquisition was started between 2 h and 2 h 30 min after spreading cells on the imaging media containing Naphthaleneacetic acid since longer induction caused meiosis I spindle collapse during the incubation period.
Condensin depletion was performed using the cut14-so strain previously described (Kakui et al., 2017 (link)) that combines transcriptional repression of cut14 under the nmt81 thiamine-repressible promoter and Cut14 depletion using an Auxin-Inducible Degron (AID) system. To induce condensin depletion, the imaging media was supplemented with 0.5 mM of 1-Naphthaleneacetic acid (Sigma) and 5 μg/ml of thiamine. For condensin depletion in meiosis I, image acquisition was started 3 h after spreading the cells on the imaging media containing Naphthaleneacetic acid. For condensin depletion in meiosis II, image acquisition was started between 2 h and 2 h 30 min after spreading cells on the imaging media containing Naphthaleneacetic acid since longer induction caused meiosis I spindle collapse during the incubation period.
7
Yeast Drug Treatment Protocols
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Yeast drug treatments were performed in YPD media at the following concentrations: 400 μg/ml 1,10 phenanthroline monohydrate (Sigma 161-0158, dissolved in ethanol), 10 μg/ml thiolutin (Santa Cruz SC-200387, dissolved in DMSO), 1 mM 1-naphthalene acetic acid (Sigma N0640, dissolved in 85% ethanol), 20 μg/ml doxycycline (Sigma D9891, dissolved in 50% ethanol), 5 μM 1-Naphthyl PP1 (Sigma CAS 221243-82-9, dissolved in DMSO), 25 μM trichostatin A (dissolved in DMSO), 10 μM α factor (Sigma custom synthesized peptide, WHWLQLKPGQPMY, dissolved in 100 mM sodium acetate, pH=5.2). mESCs were treated with Actinomycin D at 25 μg/ml (Sigma CAS 50-76-0, dissolved in DMSO).
8
Yeast Cell Culture with Auxin
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Yeast cells were grown in rich (yeast extract/peptone) or synthetic (yeast nitrogenous base with amino acid selection of plasmid maintenance) medium with 2% dextrose. For experiments using auxin in liquid cultures, 3-indoleacetic acid (I2886; Sigma-Aldrich, St. Louis, MO) was added to synthetic media to a final concentration of 500 μm. For experiments using auxin on solid media, 1-naphthaleneacetic acid (NAA; N0640; Sigma-Aldrich) was added to synthetic media to a final concentration of 1 mM.
9
Degron-Mediated Depletion of Abo1 Protein
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The insertion of the 2xHA-aid tag at the 3′ end of abo1 CDS (construct called abo1-aid) was performed in cells expressing S. pombe Skp1 in fusion to the A thaliana and Oryza sativa TIR1 F-box proteins (Egan et al, 2014 (link)). To ensure an efficient degron-dependent knock-down of the Abo1-2xHA-AID protein, all cultures were performed at 25°C, in YEA (Fig 2C ) or EMMC (Figs 2B and S4 ) (MP Biomedicals) medium. After growing cells to saturation, cell cultures were diluted to an OD600 of 0.025–0.1 in fresh medium containing 1-naphthaleneacetic acid (from Sigma-Aldrich) to a final concentration of 0.5 mM. Control cultures were treated with an identical volume of DMSO (NAA vehicle). Western blot with an antibody against anti-HA tag (Ab9110) was performed on total protein extract from cells cultivated for 6 h in the presence of NAA to verify the degradation of Abo1-2xHA-AID protein.
10
Rhizogenesis Induction in Mesembryanthemum
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To induce rhizogenesis, the explants were placed horizontally on a root-inducing medium (RIM) composed of a solid MS basal medium supplemented with 1 mg l−1 1-naphthaleneacetic acid (Sigma-Aldrich, Poland), pH 5.7. Some explants were put on this medium supplemented with 31.4 mg l−1 diphenylene iodonium (DPI, Sigma-Aldrich, Poland)—an irreversible inhibitor of flavin adenine dinucleotide (FAD) cofactor containing enzymes such as NOX (Gapper and Dolan 2006 (link)). Hypocotyls excised from M. crystallinum seedlings (day 0) and cultured for 3, 5, 7, and 10 days on RIM and RIM + DPI were used as experimental material. For each analysis, 30 hypocotyls were collected from three Petri dishes containing ten hypocotyls each.
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