EMBO/soda-nel huggingface dataset:
BAM15 Mediated Mitochondrial Uncoupling Protects Against Obesity and Improves Glycemic Control (2020)
Figure 1

Figure 1. BAM15 improves cellular respiratory kinetics by sustained mitochondrial uncoupling.

(A) Caspase 3/7 activation ± 16-hour treatment with varying concentrations of BAM15, DNP, or FCCP (N=3 per condition). 5 μM: BAM15 vs. DNP (P=0.030). 10 μM: BAM15 vs. DNP (P=0.013). 20 μM: BAM15 vs. DNP (P=0.0005), BAM15 vs. FCCP (P=0.036). 40 μM: BAM15 vs. DNP (P=0.018), BAM15 vs. FCCP (P=0.021). 80 μM: BAM15 vs. DNP (P=0.018), DNP vs. FCCP (P=0.004).

(B-F) (B) The rate of oxygen consumption (OCR), (C) ratio of oxygen consumption to extracellular acidification (OCR/ECAR), (D) maximal rate of OCR (BAM15 vs. DNP, P<0.0001; DNP vs. FCCP (P<0.0001) (E) time to maximal uncoupling rate (BAM15 vs. DNP, P<0.0001; BAM15 vs. FCCP (P<0.0001), and (F) mitochondrial respiratory half-life following acute injection of 1 μM BAM15, DNP, or FCCP (BAM15 vs. DNP, P<0.0001; BAM15 vs. FCCP (P<0.0001) (N=6 per condition).

(G, H) (G) Representative respirometry plot of intact cells ± 16-hour treatment with 20 μM BAM15 sequentially injected with oligomycin, FCCP, and a cocktail of rotenone and antimycin a (N=5 per condition) and (H) basal (P=0.002) and ATP-linked respiration, proton leak (P=0.0003), maximal uncoupling, spare respiratory capacity (P<0.0001), and non-mitochondrial respiration (N=5 per condition).

(I) Assessment of routine (R) oxygen flux (P=0.030), leak (L) (P=0.010), OXPHOS (P) and ETC (E) capacity in permeabilized C2C12 cells ± 16-hour treatment with BAM15 (N=4 per condition). PM: pyruvate and malate, ADP: adenosine diphosphate, GS: glutamate and succinate, Tm: tetramethyl-p-phenylenediamine.

(J) Citrate synthase activity ± 16-hour treatment with 20 μM BAM15 (N=10 per condition).

(K) mtDNA (COX2:18S) ± 16-hour treatment with 20 μM BAM15 (N=6 per condition).

Data information. Data are shown as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.01, ****p<0.001. Panels A, H, and I were assessed by two-way repeated measures ANOVA with Tukey's multiple comparisons. Panels D, E, and F were assessed by one-way ANOVA with Tukey's multiple comparisons. Panels J and K were assessed by unpaired Student's t-test.
BAM15 Mediated Mitochondrial Uncoupling Protects Against Obesity and Improves Glycemic Control (2020)
Figure 7

Figure 7. Increased skeletal muscle fatty acid oxidation is supported by AMPK mediated suppression of lipogenesis in white adipose tissue following treatment with BAM15.

A-D (A) Plasma concentrations of non-esterified fatty acids (NEFA) (CTRL N=7, BAM15 N=8, P=0.0073), (B) leptin (N=8 per group, P<0.0001), (C) GDF15 (N=7 per group, P=0.035), and (D) FGF21 (N=7 per group, P=0.0026) after 3-weeks of CTRL or BAM15 treatment.

E, F (E) Total (basal: P=0.015) and (F) complete (basal: P=0.0137) oxidation of 200 μM [1-14C]palmitate to CO2 from mixed gastrocnemius muscle homogenates (N=7 per group).

(G) Routine (R) oxygen flux, leak (L), OXPHOS (P) and ETC (E) capacity in permeabilized mixed gastrocnemius fibers (N=6 per group). PM: pyruvate and malate, ADP: adenosine diphosphate, GS: glutamate and succinate, Tm: tetramethyl-p-phenylenediamine, PalM: palmitoylcarnitine and malate, Oct: octanoylcarnitine, and Dur: duroquinol.

H-K (H) Representative immunoblots of phosphorylated and total AMPK and ACC from iWAT, and mixed gastrocnemius muscle and (I-K) Densitometric quantification (N=4 per group; iWAT p-AMPKT172: P=0.0186, iWAT p-ACCS79: P=0.0083, mixed gastrocnemius p-AMPKT172: P=0.0286).

(L-N) mRNA expression of transcriptional regulators of lipogenesis and lipolysis in iWAT (N=7 per group). Srebf1 (P<0.0001), Mlxipl (P=0.0018), Scd1 (P<0.0001), FASN (P=0.0009), LIPE (P=0.0015), Pnpla2 (P<0.0001), PPARg (P<0.0001).

Data information: Data are shown as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.01, ****p<0.001. Panels E, F, and G were assessed by two-way repeated measures ANOVA with Tukey's multiple comparisons. Panels A, B, C, D, I, J, K, L, M, and N were assessed by an unpaired Student's t-test.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 1

Figure 1. Accumulation of Gb3 in Batten Disease cellular models.

(A) Representative Opera images of WT and CLN3 KO ARPE 19 cell lines stained using fluorescent-conjugated cholera toxin (to detect GM1), LipidTox (to detect neutral lipids), fluorescent-conjugated Shiga toxin and is quantification (to detect Gb3), Filipin III (to detect cholesterol). Data are presented as mean ± SD ***: P ≤ 0.0001, as determined by Student's t‐test (n=3 biological replicas in duplicate). Scale bars: 50 µm.

(B-C) Representative confocal images and their quantification of Gb3 accumulation within the lysosome, detected by fluorescent-conjugated Shiga toxin, from WT, CLN3-KO and CLN7-KO ARPE-19 cells treated with PDMP or silenced for Gb3 synthase (siGb3S) (*** vs WT, °°° vs DMSO). Data are presented as mean ± SD, °°°/***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

(D) Lipidomic analysis of ARPE-19CLN3KO and ARPE-19CLN7KO. Data are presented as mean ± SD, *: P ≤ 0.01, **: P ≤ 0.001, ***: P ≤ 0.0001, as determined by ANOVA (n=4 technical replicas).
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 2

Figure 2. Neural accumulation of Gb3 in brain areas of the CLN7Δex2 mouse.

(A-B) Representative confocal images of Gb3 accumulation, revealed by STX staining, in brain sections from CLN7Δex2 mice at 3 and 7.5 months of age compared with CLN7 WT mice. Scale bars: 80 µm.

(C-D) Gb3, NeuN, and GFAP distribution in brain areas of CLN7Δex2 mice at 7.5 months of age. Scale bars: 40 µm, 20 µm.

(E) Lipidomic analysis of neurons isolated from CLN7 WT and CLN7Δex2 mouse forebrain. Data are presented as mean ± SD, **: P ≤ 0.001, ***: P ≤ 0.0001, as determined by ANOVA (n= 3 biological replicas)
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 3

Figure 3. Role of Gb3 in Batten Disease and identification of correctors of Gb3 accumulation.

(A) Representative confocal images and quantification of SCMAS staining within the lysosome of ARPE-19 WT and CLN3 KO upon depletion of Gb3S (siGb3S), LacCer (siLCS), and GM3s (siGM3S). (*** vs WT, °°° vs CLN3 KO siSCR). Data are presented as mean ± SD, °°°/***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

(B) Identification of FDA-compounds reducing Gb3 accumulation: Plot showing the ability of compound hits to reduce Gb3 within the lysosomal compartment compared with DMSO treated mutant cells. Data are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate).

(C) Table showing the EC50s (Half maximal effective concentration) from the dose-response curves of compound hits. EC50s were calculated using the Prism software.

(D) Representative confocal images and quantification of STX staining within the lysosome of ARPE-19 WT and CLN7 KO in DMSO or treated 48h with Tamoxifen. (*** vs WT, °°° vs CLN3 KO DMSO). Data are presented as mean ± SD, °°°/***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

(E) Representative confocal images and quantification of STX staining within the lysosome of NPCs WT and derived from CLN7Pa474 patient IPSCs in DMSO or treated with Tamoxifen for 48h. (*** vs WT, °°° vs CLN7Pa474 DMSO). Data are presented as mean ± SD, °°°/***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 4

Figure 4. Tamoxifen-mediated clearance of Gb3 is ER-independent but TFEB-dependent.

(A) Representative confocal images and Quantification of STX within the lysosome in U2-OS and HeLa cells after acute silencing of CLN3 (siCLN3) in DMSO or treated 48h with Tamoxifen. (*** vs siSCR DMSO, °°° vs siCLN3 DMSO). Data are presented as mean ± SD, °°°/***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

(B) Representative confocal image and quantification of Gb3 in ARPE-19 CLN3 KO cells silenced with siRNA against scramble sequence and TFEB (siTFEB) for 72h and treated for the last 48h with DMSO or Tamoxifen. Data are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

(C) Representative confocal image and quantification of TFEB in U2-OS cells silenced with siRNA against scramble sequence and CLN3 (siCLN3) for 72h and treated for the last 48h with DMSO or Tamoxifen (5 µM and 10 µM). Data are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 5

Figure 5. Tamoxifen dephosphorylates TFEB.

(A) Immunoblot analysis and quantification of pTFEB S211 in HeLa cells stably expressing TFEB-GFP. Data are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas ). GAPDH immunoblotting was performed as a loading control.

(B) Immunoblot analysis of TFEB shift in ARPE-19 CLN3 KO cells. β-actin immunoblotting were performed as a loading control.

(C) Representative confocal image and quantification of TFEB localization in HeLa TFEB-GFP transfected with RagC for 48h and treated for the last 3h with DMSO, Torin1, Tamoxifen or Ospemifene. Ratios of nuclear to cytosolic TFEB localization in RagC non-expressing (RagC-) and RagC-expressing cells (RagC+) are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.

D) Representative confocal images and quantification of STX in ARPE-19 CLN3 KO cells transfected with empty vector or HA-RagC for 48h and treated for 48h with DMSO or Tamoxifen. STX average spot area presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (n=3 biological replicas in duplicate). Scale bars: 20 µm.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 6

Figure 6. Tamoxifen reduce Gb3 accumulation in brain of CLN7Δex2 mice.

Representative confocal images of STX in the Cortex, Hippocampus, and Cerebellum brain section derived from 7.5 months-old mouse WT or CLN7Δex2 injected with vehicle or Tamoxifen (Tamox). Quantification of confocal images, the plot shows the quantification of the STX average spot area normalized for the number of Hoechst positive cells. (*** vs WT, °°°/°° vs CLN7Δex2 Vehicle). Data are presented as mean ± SD, **/oo: P ≤ 0.001, ***/ooo: P ≤ 0.0001, as determined by ANOVA (N≥3 biological replicas). Scale bars: 60 µm.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 7

Figure 7. Tamoxifen reduces SCMAS accumulation in the brains of CLN7Δex2 mice.

Representative confocal images of SCMAS in the Cortex, Hippocampus, and Cerebellum brain section derived from mouse WT or CLN7Δex2 injected with vehicle or Tamoxifen. Quantification of confocal images, the plot shows the quantification of the SCMAS average spot area normalized for the number of Hoechst positive cells. (*** vs WT, °°° vs CLN7Δex2 Vehicle). Data are presented as mean ± SD, ***: P ≤ 0.0001, as determined by ANOVA (N≥3 biological). Scale bars: 60 µm.
Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype (2021)
Figure 8

Figure 8. Tamoxifen ameliorates CLN7Δex2 phenotype.

(A-B) Representative confocal images and quantification of IBA-1 in the Cortex, Hippocampus, and Cerebellum brain section derived from WT or CLN7Δex2 mice injected with the vehicle or Tamoxifen. (***/**/* vs WT, °°°/°°/° vs CLN7Δex2 Vehicle). Data are presented as mean ± SD, */o: P ≤ 0.01, **/oo: P ≤ 0.001, ***: P ≤ 0.0001, as determined by ANOVA (N≥3 biological replicas). Scale bars: 50 µm.

(C) Plots show the quantification latency to fall from the rotarod. Data are presented as mean ± SD, *: P ≤ 0.01, as determined by ANOVA (N≥3 biological replicas).

(D) Representative Images of hindlimb clasping test in mouse WT or CLN7Δex2 injected with vehicle or Tamoxifen.

Expanded View Legends
WRNIP1 protects stalled forks from degradation and promotes fork restart after replication stress (2016)
Figure 1

Figure 1. Loss of WRNIP1 leads to nascent DNA strand degradation after HU-induced replication stress

(A) Western blot analysis showing the expression of the WRNIP1 protein in wild-type cells (shWRNIP1WT) and WRNIP1-deficient (shWRNIP1) or mutant (shWRNIP1T294A) cells. MRC5SV fibroblasts were used as a positive control. The membrane was probed with an anti-FLAG or anti-WRNIP1. GAPDH was used as a loading control. Below each lane of the blot the ratio of WRNIP1 protein to total protein, then normalised to MRC5SV, is reported.

(B) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were pulse-labelled with CldU, and then subjected to a pulse-labelling with IdU.

(C) Analysis of replication fork velocity (fork speed) in the cells under unperturbed conditions. The length of the green tracks were measured. Mean values are represented as horizontal black lines (ns, not significant; Student’s t-test).

(D) Cells were treated as in (B). For each replication origin, the length of the right-fork signal was measured and plotted against the length of the left-fork signal. A schematic representation of symmetric and asymmetric forks is given. If the ratio between the left-fork length and the right-fork length deviated by more than 33% from 1 (that is, outside the violet dashed lines in the graphs), the fork was considered asymmetric. The percentage of asymmetric forks was calculated for all cell lines. N= number of forks counted for each cell line. R represents linear correlation coefficient.

(E) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were pulse-labelled with CldU, treated with 4mM HU and then subjected to a pulse-labelling with IdU.

(F) Graphs show the percentage of red (CldU) tracts (stalled forks) or red-green (CldU-IdU) contiguous tracts (restarting forks) in the cells. Mean shown, n = 3. Error bars represent standard error. (*, p < 0.05; **, p < 0.01; Student’s t test). Representative DNA fiber images are shown. Scale bars, 10 µm.

(G) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were sequentially pulse-labelled with CldU and IdU as indicated, then treated or not with 4 mM HU.

(H) Representative IdU tract length distributions in all cell lines under unperturbed conditions (left graph) or after HU treatment (right graph). Median tract lengths are given in parentheses. See also Tables S1 and S2 for details on the data sets and statistical test. Representative DNA fiber images are shown. Scale bars, 10 µm.
WRNIP1 protects stalled forks from degradation and promotes fork restart after replication stress (2016)
Figure 2

Figure 2. Inhibition of MRE11 exonuclease activity prevents nascent DNA strand degradation after replication stress

(A) Experimental scheme of dual labelling of DNA fibers in wild-type cells (shWRNIP1WT) or WRNIP1-deficient cells (shWRNIP1). Cells were sequentially pulse-labelled with CldU and IdU as indicated, then left untreated or treated with 4 mM HU in combination or not with 50 µM Mirin.

(B) Representative IdU tract length distributions in shWRNIP1WT (left graph) or shWRNIP1 cells (right graph) after treatment. Median tract lengths are given in parentheses. See Tables S1 and S2 for details on the data sets and statistical test. Representative DNA fiber images are shown. Scale bars, 10 µm.
WRNIP1 protects stalled forks from degradation and promotes fork restart after replication stress (2016)
Figure 4

Figure 4. RAD51 protects nascent DNA strand from degradation after fork stalling in the absence of WRNIP1

(A) Experimental scheme of pulse-labelling of DNA fibers in wild-type cells (shWRNIP1WT) or WRNIP1-deficient cells (shWRNIP1). Cells were labelled with IdU and exposed or not to 25 μM RAD51 inhibitor, then treated or not with 4 mM HU.

(B) Representative IdU tract length distributions in shWRNIP1WT cells (left graph) or shWRNIP1 cells (right graph). Median tract lengths are reported in parentheses. See Tables S1 and S2 for details on the data sets and statistical test. Representative DNA fiber images are reported. Scale bars, 10 µm.

(C) Scheme of DNA fiber tract analysis in shWRNIP1 cells. Cells were transfected with an empty vector or a plasmid expressing a wild-type human RAD51, and 48 h thereafter labelled with IdU and treated or not with 4 mM HU.

(D) Representative IdU tract length distributions in shWRNIP1 cells or shWRNIP1 cells expressing exogenous wild-type RAD51 after HU exposure. Median tract lengths are given in parentheses. See Tables S1 and S2 for details on the data sets and statistical test. Representative DNA fiber images are given. Scale bars, 10 µm. Western blot shows the expression of the RAD51 protein in shWRNIP1 cells. The membrane was probed with an anti-RAD51. LAMIN B1 was used as a loading control.
ABCB-mediated shootward auxin transport feeds into the root clock (2023)
Figure 2

Figure 2. The amiR-2572 line has reduced auxin oscillation amplitude and LR density defects.

A Luciferase image of the whole WT and amiR-2572 seedlings after 10 min exposure to visualize the pre-branch sites in the root by DR5:LUC luminescence.

B Quantification of pre-branch site density in 10-day-old WT and amiR-2572 determined by DR5:LUC luminescence, n= 26 (WT and amiR-2572).

C Confocal images of root tips in 5-day-old Col-0, amiR-2572 and syn-tasi-1522A#1 seedlings. LRC cells were indicated by white asterisks. The distal LRC cells were indicated by white arrows The scale bar represents 50 µm.

D-F Quantification of meristem size (D), LRC cell number (E), and distal LRC cell length (F) in 5-day-old Col-0, amiR-2572 and syn-tasi-1522A#1 seedlings. The meristem size was measured along the yellow dashed lines in (C), as estimated by the distance from the QC to the first elongating cortical cell, nD/E/F= 33/30/22 (WT), 34/30/26 (amiR-2572) and 37/30/23 (syn-tasi-1522) from 3 independent biological repeats.

G Macroview stereo microscopic view of DR5:VENUS expression in root tips of 3-day-old WT, amiR-2572 and syn-tasi-1522A#1 seedlings, red arrows indicate DR5 stripes in the lateral root cap. Scale bar = 50 µm

H Quantifications of the time interval(s) between the consecutive disappearance of DR5:VENUS stripes in the most-distal lateral root cap in 3-day-old WT, amiR-2572 and syn-tasi-1522A#1. n= 15 (WT), 18 (amiR-2572) and 17 (syn-tasi-1522A#1)

I, J Quantification of the oscillation period (I) and amplitude (J) of DR5::LUC in 3-day-old WT and amiR-2572, n= 21 (WT and amiR-2572).

Data information: For (B, I, J), Unpaired two-tailed Student's t-test with Welch's correction, P<0.05 (*), P<0.001(***). For (D-F, H), One-way ANOVA in combination with Tukey's multiple comparisons test, significant differences (P ≤ 0.05) are indicated by different lowercase letters. Central bands in the box plots show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles, outliers are represented by dots.
ABCB-mediated shootward auxin transport feeds into the root clock (2023)
Figure 3

Figure 3. Analysis of plasma membrane localisation and cellular transport activities.

A Co-localisation of YFP-ABCB15 with Propidium Iodide (PI) in root epidermal cells of 3-day-old seedlings. A plot of the fluorescence intensity of YFP and PI along the dashed lines shows the colocalization of the YFP and PI intensity peaks. Scale bars = 10 µm.

B Presence of YFP-ABCB15 on Hechtian strands of epidermal cells of 3-day-old seedlings after 10min 0.8M mannitol treatment. Cell walls are counterstained by Propidium Iodide (PI). The white arrow indicated the Hechtian strands. Scale bars = 10 µm.

C IAA export assay. Export of [3H]-IAA, assayed in parallel from tobacco mesophyll protoplasts expressing ABCB1, ABCB15-22 and ABCB17P980G against vector control. mean ± SE; n = 26 (vector control), 44 (ABCB1), 5 (ABCB15), 4 (ABCB16), 9 (ABCB17), 5 (ABCB18), 9 (ABCB22) and 8 (ABCB17P980G),transport experiments generated from independent tobacco transfections.

D Export assay of plant hormones IAA (nVC/ABCB17 = 26/9), IBA (nVC/ABCB17 =6/6), BA (nVC/ABCB17 =7/7), ABA (nVC/ABCB17 =6/6), tZ (nVC/ABCB17 =7/6) and malate (nVC/ABCB17 =6/16) in parallel from N. benthamiana mesophyll protoplasts expressing ABCB17 against vector control. mean ± SE; transport experiments generated from independent tobacco transfections.

E [3H]-IAA export from WT, abcb15-1, abcb16-1, abcb17-1, abcb18-1, abcb22-1, amiR-2572 and syn-tasi-1522A#1 Arabidopsis leaf mesophyll protoplasts, mean ± SE; n= 9 (WT), 4 (abcb15-1), 3 (abcb16-, abcb17-1, abcb18-1) and 4 (abcb22-1), n = 4 (WT, amiR-2572) and 6 (syn-tasi-1522A#1)

Data information: For (C-E), Unpaired two-tailed Student's t-test with Welch's correction, P<0.05 (*), P<0.01 (**).
ABCB-mediated shootward auxin transport feeds into the root clock (2023)
Figure 4

Figure 4. ABCB15-22 contribute to shootward auxin transport for LR induction.

A Shootward auxin transport assay of [3H]-IAA and [14C]-BA in WT (Col-0) and amiR-2572 roots, mean ± SE; n = 3 (Col-0 and amiR-2572; IAA); n=4 (Col-0; BA) and 3 (amiR-2572; BA)

B Analysis of DR5::VENUS expression in the root elongation zone of 4-day-old pWOX5:XVE>>YUC1-2A-TAA1, in Control and amiR-2572 treated with β-estradiol (5 µM) for 0, 7.5 and 9h. Images are composed of several tiles generated in a single snap with automatic assembly, PI in gray. The zoomed images of the yellow squares are presented below each root showing the accumulation of the DR5::VENUS signal in the elongation zone. Scale bar = 100µm.

C Quantification of DR5::VENUS signals in the epidermis of the elongation zone shown in (B). n0h=40/40, n7.5h=70/70, n9h=140/160 (WT/amiR-2572) from at least 4 (0h), 7 (7.5h), 14 (9h) seedlings of each treatment

D LR density of pWOX5:XVE>>YUC1-2A-TAA1, in control and amiR-2572 treated with β-estradiol. 7-day-old seedlings were transferred to MS plates containing 500 nM β-estradiol. The primary root length and the total number of emerged LRs were recorded after 5 and 8 days of treatment.n = 9 (Col-0; Mock), 7 (amiR-2572; Mock), 11 (Col-0; Estradiol) and 10 ( amiR-2572; Estradiol)

Data information: For (A, C, D), Unpaired two-tailed Student's t-test with Welch's correction, P<0.05 (*), P<0.001 (***). Central bands in the box plots show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles, outliers are represented by dots.
ABCB-mediated shootward auxin transport feeds into the root clock (2023)
Figure 5

Figure 5. ABCB15-22 expressed in the outer layers of the root meristem.

A, B Expression pattern of proABCB15:NLS-GFP-GUS, proABCB16:NLS-GFP-GUS, proABCB17:NLS-GFP-GUS, proABCB18:NLS-GFP-GUS, and proABCB22:NLS-GFP-GUS expression in roots of 3-day-old seedlings, using confocal microscopy showing longitudinal overview pictures (Propidium iodide in magenta), and images zoomed on the epidermis and LRC corresponding to yellow squares (A); cytological sections of GUS stained roots of 3-day-old seedlings, counterstained with ruthenium red (B). Scale bars represent 20 µm for both graphs.

C Root annotation schematic representation of the summary expression pattern of ABCB15-22 in the root meristem.