EMBO/SourceData huggingface dataset:
Figure 3A (Left) The representative cross-section image showing nuclear distribution of MATR3 and H3K27me3 before and after 12h treating with antisense L1 ASOs in AML12 cells. (Right) Line charts showing pixel intensity of each channel on the ROIs. r, coefficient of correlation. Scale bars, 5μm.C (Left) The representative cross-section image showing nuclear distribution of H3K27me3 and AS L1 RNA before and after MATR3 knockdown (Dox treatment for 3d) in AML12 cells. (Right) The normal distribution curve for the AS L1 pixel intensity. Scale bars, 5μm.E Representative images showing nuclear colocalization of AS L1 RNAs with wild-type and truncated GFP-MATR3 proteins in AML12 cells. Scale bars, 5μm.G Representative cross-section images showing nuclear localization of H3K27me3 upon Ctrl, MATR3 knockdown and exogenous GFP-MATR3 (WT/△ZF1/△ZF2/△RRM1/△RRM2) overexpression in AML12 cells. Scale bars, 5μm.
Figure 4A Representative images of the GFP-MATR3 (WT/△RRM2) FRAP experiments in AML2 cells. Solid arrows indicate the bleached points. Scale bars, 5μm.C Representative images of droplet formation assays with different concentrations of GFP-MATR3 proteins. NaCl concentration, 50mM. Scale bars, 5μm. D Representative images of droplet formation assays with different NaCl concentrations. GFP-MATR3 protein concentration, 3 μM. Scale bars, 5μm. E Areas of MATR3 protein droplets formed in different protein concentration (3 μM: n = 315; 2 μM: n = 196; 1 μM: n = 165; 0.5 μM: n = 50). The P values were calculated using unpaired two-tailed Student's t test; ****p<0.0001. F Areas of MATR3 protein droplets formed in different NaCl concentration (50 mM: n = 248; 100 mM: n = 196; 400 mM: n = 127; 800 mM: none). The P values were calculated using unpaired two-tailed Student's t test; ****p<0.0001.H Representative images of droplet formation assays by GFP-MATR3 with different concentration (0nM, 5nM, 50nM, 200nM, 500nM) of AS L1 RNAs. GFP-MATR3 protein concentration, 3 μM. NaCl concentration, 50mM. Scale bars, 5μm.
Figure 5E Snapshot of an example region, showing Hi-C, H3K27me3 ChIP-seq and AS L1 in MATR3 RIP-seq in Ctrl and/or shMatr3 samples, using HiCExplorer. The values on the y-axis for Hi-C contact and O/E heatmap are iced normalized read counts at 100kb resolution. The values on the y-axis for ChIP-seq and RIP-seq are average reads per million of mapped reads (RPM).K Percentage of TADs located in compartment A or B regions from five TAD groups (same groups in J).L Boxplot shows gene expression changes for TAD groups from J. The number of genes in each groups are 2050, 2094, 2356, 2071 and 1605. The box of the boxplot covers the 50% of the values between 1st quartile and 3rd quartile of each group; Boxplot whiskers represent the minimal and maximal values of each group; The horizontal line of box represent the median value of each group; Red dashed lines indicate the zero.
Figure 6A Representative Western blot on chromatin fractionation experiments of Ctrl and shMatr3 cells. Each fraction was loaded with same number of cells. GAPDH, histone H3, Lamin A/C were used as positive controls for S1, S2 and S3, S4, respectively.B Genome browser view of the SAMMY-Seq and H3K27me3 ChIP-seq in Ctrl and shMatr3 (replicate 1 and 2 merged) cells at a representative region. Gray boxes under each SAMMY-seq track showing the SAMMY-seq domains. The shadows showing the shMatr3 loss SAMMY-seq domains.H Average MATR3 RIP-seq sense and antisense signal at L1 loci around H3K27me3 domains (groups as in F) with 200kb upstream and downstream flanking regions from AML12 cells.
Figure 7C MATR3 knockdown and GFP-tagged WT/S85C/F115C MATR3 protein replacement in N2A cells. The efficiency of endogenous MATR3 knockdown and exogenous GFP-MATR3 (WT/S85C/F115C) over-expression as detected by western blotting. Representative of two independent replicates with similar results.D Representative images of the GFP-MATR3 (WT/S85C/F115C) FRAP experiments in N2A cells. Solid arrows point to the bleached points. Scale bars, 5μm.G Representative cross-section images showing nuclear localization of H3K27me3 upon Ctrl, MATR3 knockdown and exogenous MATR3 (WT/S85C/F115C) overexpression in N2A cells. Scale bars, 5μm. H Standard deviation of H3K27me3 pixel intensity upon Ctrl (n=93), MATR3 knock down (n=100), GFP-MATR3 (WT) rescue (n=104), GFP-MATR3 (S85C) rescue (n=82) and GFP-MATR3 (F115C) rescue (n=120). The P values were calculated using unpaired two-tailed Student's t test; ns, not significant, *p<0.05, ****p<0.0001. Error bars indicate mean ± s.e.m.
Figure 2U2Os cells co-transfected with YFP-FKBP and PCM1F2-Ce3-FRB, PACT-Ce3-FRB, or Ce3-FRB-Centrin2 were treated with rapamycin (100 nM). The translocation of YFP-FKBP onto PCM1F2- Ce3-FRB-labeled centriolar satellites, the PACT-Ce3-FRB-labeled pericentriolar matrix, and the Ce3-FRB-Centrin2-labeled centriolar lumen was monitored. The normalized fluorescence intensity of YFP-FKBP accumulation at centrosomes upon rapamycin (100 nM; blue) and DMSO (0.1%, vehicle control; red) treatment. Data are shown as the mean ± S.E.M. The graphs show immediate translocation of YFP-FKBP after rapamycin induction at centriolar satellites (left, n = 24 cells), the pericentriolar matrix (middle, n = 20 cells), and the centriolar lumen (right, n = 23 cells) from seven independent experiments.
Figure 3U2Os cells co-transfected with each YFP-FKBP-labeled varisized probe (YFP-FKBP, YFP-FKBP-Grp1, YFP-FKBP-Luciferase, YFP-PSD95-FKBP, YFP-FKBP-β-Gals, YFP-FKBP-β-Galm, YFP-FKBP-ΔNβ-Gal, and YFP-FKBP-β-Gal) and Ce3-FRB with labels specific for centrosomal sub-compartments were treated with rapamycin (100 nM). The images of cerulean channel show centrosome sub-compartment from the untreated cells. Arrowheads indicate sites of centrosomes. Insets show higher-magnification images of the centrosomal regions. Dotted lines indicate the cell boundaries. The normalized fluorescence intensity of each probe at centriolar satellites (left, n = 14, 19, 17, 13, 11, 22, 14, and 9 cells from small to large probes), pericentriolar matrix (middle, n = 20, 19, 18, 16, 10, 18, 10, and 12 cells from small to large probes), and centriolar lumen (right, n = 10, 13, 13, 6, 3, 19, 13, and 9 cells from small to large probes) from 3-6 independent experiments.
Figure 4Depolymerization of microtubules by Nocodazole (10 µM) treatment. U2Os cells that expressed PACT-Ce3-FRB (blue, a marker of centrosomes) were incubated with antibodies against α-tubulin to label microtubules (red) in the absence (0.1% DMSO) or presence of Nocodazole treatment for 20 min. Lower panels show magnified images of the centrosomal regions. The normalized intensity of α-tubulin around centrosomes in the absence or presence of Nocodazole treatment for 20 min. The circular ROIs in diameter of 3 µm, centering PACT-labeled centrosomes, were used for measurement of the level of microtubule around centrosomes. Data (black) are shown as the mean ± SD. (n = 196 and 156 cells from left to right; 3 independent experiments).U2Os cells expressing PACT-Ce3-FRB (blue, a marker of centrosomes) were incubated with phalloidin to label the F-actin (red) with DMSO (0.1%), CK666 (0.4 mM) for 2 hr, or Latrunculin A (LatA, 0.5 mM) for 1 hr. Lower panels show the magnified images of the centrosomal regions. The normalized intensity of actin filaments around centrosomes in the absence or presence of CK666 or LatA treatment. The circular ROIs in diameter of 3 µm, centering PACT-labeled centrosomes, were used for measurement of the level of actin filaments around centrosomes.
Figure 5HeLa cells co-transfected with H2B-RFP (a chromosome marker), PACT-Ce3-FRB, and YFP-FKBP-Luciferase were sequentially incubated with thymidine (2 mM) for 16-18 hr and RO3306 (2.5 ng/mL) for 12 hr to synchronize cells in G2/M phase (Interphase cells in left panel). The probe trapping experiment in interphase cells was performed right after RO3306 washout. Anaphase cells were chosen for probe trapping experiment 45-60 min after RO3306 washout. Transfected cells in interphase or anaphase were treated with 100 nM rapamycin (Rapa). The centrosome regions are shown in lower panels. The normalized fluorescence intensity of YFP-FKBP-Luciferase accumulation at centrosomes in interphase (Green, n = 12 cells) or anaphase (Red, n =10 cells) cells upon rapamycin (100 nM) treatment.HeLa cells co-transfected with H2B-RFP (a chromosome marker), PACT-Ce3-FRB, and YFP-FKBP-Luciferase were synchronized in anaphase with or without CK666 treatment (0.4 mM). Rapamycin (100 nM) was added for 5 min to trigger the YFP-FKBP-Luciferase trapping at centrosomes. The right panel is the enlarged images of centrosome regions and arrows indicated centrosome sites. The percentage of cells in D exhibiting probe translocation. Data are shown as mean ± S.E.M. (n = 47 and 31 cells in DMSO and CK666 treated groups, respectively; 3-4 independent experiments).
Figure 6U2Os cells were incubated with antibodies against 𝑟-tubulin (green; a marker of 𝑟-TuRC) or Pericentrin (red; a marker of centrosomes) in the absence (0.1% DMSO) or presence of CK666 treatment (0.4 mM) for 2 hr. The right panel shows magnified images of the indicated region of interest (ROI). S The normalized intensity of 𝑟-tubulin at centrosome in control (0.1% DMSO, 2 hr) or CK666 (0.4 mM, 2 hr)-treated cells. Individual data points and the mean ± SD (in red) are shown (n = 150 and 150 cells in DMSO and CK666-treated group, respectively; four independent experiments).U2Os cells transfected with GCP4-GFP were treated with or without CK666 (0.4 mM, 2 hr) and then observed for the indicated time after photobleaching. The mobile fraction percentage (left) and mobile recovery half-time (right) of GCP4- GFP in FRAP experiments (c). Data (red) are shown as meanU2Os cells expressing EB1-mNeon were incubated with 0.1% DMSO or 0.4 mM CK666 for 2 hr. Images were captured over 150 sec and were overlaid to show the overall tracks of the microtubules. The insets show enlarged images of the centrosomal regions. F, G. The number of microtubule tracks that emanate from centrosomes (F) and the microtubule elongation rate (G) in the absence and presence of CK666 treatment (0.4 mM). Individual data points and the mean ± SD (in red) are shown (n = 78449 and 54283 microtubule tracks from 27 and 22 cells in the DMSO- and CK666-treated groups, respectively; three independent experiments).
Figure 1(A) Super-resolved average intensity (bar: high 8 to low 0 density), average diffusion coefficient (bar: Log10D high 1 to low -5 mobility) and trajectory maps (16000 frames) of BoNT/Aiwt-At647N on the plasma membrane (uPAINT) and following endocytosis (sdTIM).B, C Number of BoNT/Aiwt-At647N and mutant trajectories (B) on the plasma membrane (uPAINT) and (C) following endocytosis (sdTIM). N = 17 technical replicates/ condition from 5-6 independent biological replicates(E) EM analysis of endocytosed HRP-tagged BoNT/Ai. Representative images show cytochemically stained HRP precipitate (arrowheads). Endo/lysosomes, multivesicular bodies (MVB), structures with the morphology of autophagosomes (AP), and (i) presynaptic (Ps) and (ii) axonal (Ax) plasma membrane (PM). Postsynaptic density (asterisks), non-stained endocytic compartments (arrows) and background binding on the PM (arrows in ii) are indicated. Scale bars 100 nm.F, G (F) Percentage of total BoNT/Aiwt-HRP and indicated mutants localized on the PM or in the indicated endocytic structures, and (G) the average sectional area of the HRP-stained endocytic compartments. (F,G) n = 60 electron micrographs/ condition from 3 independent biological replicates.
Figure 2A, B Low resolution images and the corresponding super-resolved uPAINT average intensity (bar: high 8 to low 0 density), average diffusion coefficient (bar: Log10D high 1 to low -5 mobility) and trajectory maps of SV2Awt-pH/At565nb in (A) control conditions (high K+; Ctrl), and (B) following treatment with 100 pM BoNT/Aiwt-At647N (supplemented in high K+).(C) Scatter plots of the average diffusion coefficient (µm2 s-1) single-molecule mobility of SV2Awt-pH/At565nb and SV2AT84A-pH/565nb in the control condition (high K+; Ctrl) and following treatment with 100 pM BoNT/Aiwt-At647N or BoNT/AiSV2-At647N (supplemented in high K+). (D) Scatter plots of the average diffusion coefficient (µm2 s-1) single-molecule mobility of Syt1wt-pH/At565nb, Syt1K362A,K328A-pH/565nb and Syt1K52A-pH/At565nb in the control condition (high K+; Ctrl) and following treatment with 100 pM BoNT/Aiwt-At647N (supplemented in high K+).
Figure 3A-C (A) Representative low-resolution image of SV2Awt-pH (green), and corresponding dual-colour super-resolved uPAINT trajectories of SV2Awt-pH/At565nb (cyan) imaged simultaneously with BoNT/Aiwt-At647N (magenta). Regions (i-ii) are magnified below, and (iii) is magnified in (B) showing a representative time-lapse sequence of the single molecules of SV2Awt-pH/At565nb (cyan) and BoNT/Aiwt-At647N (magenta) detected coincidentally in a neuronal projection. The 10 x 10 pixel boxed region (iv) is magnified in (C), showing frame-by-frame coincidental detection of BoNT/Aiwt-At647N and SV2Awt-pH/At565nb obtained using a 50 Hz (20 ms exposure time) 16,000 frame acquisition as indicated.H, I Representative images of NASTIC nanocluster analysis of SV2Awt-pH/At565nb in (H) the control condition (high K+; Ctrl) and (I) following high K+ stimulation in the presence of 100 pM BoNT/Aiwt-At647N, with 2D kernel density estimation (KDE, high density in red), instantaneous Log10 diffusion coefficient map (low mobility in red), and nanoclusters with their respective trajectories (white) and centroids (orange) shown. Boxed areas are magnified on the right, with arrowheads indicating nanoclusters.J-N Quantification of the (J) SV2Awt-pH/At565nb, (K) SV2AT84A-pH/At565nb, (L) Syt1wt-pH/At565nb, (M) Syt1K326A,K328A-pH/At565nb and (N) Syt1K52A-pH/At565nb nanoclustering in control conditions (high K+; Ctrl) and following BoNT/Aiwt-At647N treatment (diluted in high K+). (J-N) N = 11-19 technical replicates from 3-5 independent biological replicates,
Figure 4(A) Representative confocal Z-stack sum projections of neurons transduced with TagBFP2-expressing (blue) control (ctrl; non-targeting sgRNA) and CRISPRi Syt1 KD (Syt1 sgRNA1-3) lentiviruses. Neurons were treated for 30 min with 10 units of AbobotulinumtoxinA, and immunostained for endogenous Syt1 (yellow) and BoNT/A-cleaved SNAP-25 (SNAP-25/A; magenta). Boxed regions (i-iv) are magnified on the right, with neuronal somas outlined by dashed lines for clarity. B, C Quantification of mean fluorescence intensity (MFI) of endogenous (B) Syt1, and (C) cleaved SNAP-25 (SNAP-25/A), in control and Syt1 sgRNA1-3 KD neurons following AbobotulinumtoxinA treatment. (B-C) n = 21 technical replicates/ condition from 2 independent biological replicates. Statistical significance was assessed using a non-parametric Kruskal-Wallis multiple comparison testH, I Quantification of (H) Syt1 and (I) SNAP-25/A MFI in non-transduced neurons (neurons negative for TagBFP2) with and without pLenti6.3-Syt1wt-pH expression following AbobotulinumtoxinA treatment (10 units, 30 min.) n = 3 technical replicates/ condition from one biological experiment.
Figure 5(A) Representative patch-clamp spontaneous miniature inhibitory postsynaptic current (mPSC) recordings in naïve neurons, and in control (non-targeting) and Syt1 KD (CRISPRi sgRNA1) neurons following a 30 min treatment with AbobotulinumtoxinA (10 units). B, C Quantification of mPSC (B) peak amplitudes and (C) frequencies for indicated conditions. Error bars are shown as standard error of the mean (±SEM). N = 5-12 technical replicates/ condition from 2 independent biological replicates in (B, C) statistical significance was assessed using a non-parametric Kruskal-Wallis multiple comparison test (B), an ordinary one-way ANOVA multiple comparison test (C)(D) EM analysis of endocytosed BoNT/Aiwt-HRP following lentiviral control (ctrl; non-targeting sgRNA) and Syt1 sgRNA1 CRISPRi KD transduction. HRP precipitate in synaptic vesicles (SVs; arrowheads) and tubular structures (arrows) in presynapses (Ps) are indicated. The postsynaptic density is marked with an asterisk. (E) Percentage of total BoNT/Aiwt-HRP localized in SVs, endo/lysosomes (E), tubular structures (T), multivesicular bodies (MVB), structures resembling autophagosomes (AP) and on the plasma membrane (PM) in the indicated conditions. n = 56 regions of interest/ condition from 2 independent biological replicates (E) two-way ANOVA multiple comparison test (E).
Figure 1C. Dissociation constant values for wild-type AtAGO10, HsAGO2, and the AtAGO10+Hs-loop HsAGO2+At-loop mutants binding to a target RNA with base pairing complementarity to the miRNA seed region (guide nucleotides 2-7). Guide-target base pairing schematic is shown above. Plotted data represent the best-fit KD values from n=3 technical experimental trials, each containing 12 data points. Error bars indicate s.e.m. ANOVA with Dunnett's posthoc test P values: **** < 0.0001. No data points were excluded from the analyses.
Figure 2C. Dissociation constants derived from data in panel B. Data points in represent best-fit KDE. Fraction of target RNA released in the fast phase of curve fits in panel D. Data points in E represent best-fit KD
Figure 5B. Dissociation constants derived from data in panel A. Data in panels B represent best-fit values of data in panels No data points were excluded from the analyses.D. Concentration of cleaved product generated by HsAGO2 or HsAGO2+At-loop at 22 ˚C or 36 ˚C versus time. were obtained in n=3 independent technical trials. No data points were excluded from the analyses.E. k2 and k3 values derived from fitting data in D to the burst-and-steady-state equation Data in panels E represent best-fit values of data in panels No data points were excluded from the analyses.
Figure 6B. Fold repression of Renilla expression (compared to reporter plasmid transfection alone) versus the amount of transfected siRNA duplex, wild-type HsAGO2-siRNA complex, or HsAGO2+At-loop-siRNA complex. The 2-8 target (left panel) indicates miRNA-like targeting. The 2-19 target (right panel) indicates siRNA-like targeting. Dotted line indicates the normalized expression level of the positive control (no repression). Graphed data in B represent the individual and the ratio of mean values of n=3 independent biological trials, respectively. Error bars indicate s.e.m. ANOVA with Dunnett's posthoc test P values: ** = 0.0062, * = 0.0363. No data points were excluded from the analyses.
Figure 1B. Numbers of CD4+ and CD8+ in 1×106 uterine cells on gd 18.5 after preconceptual infection of the mother in comparison with non-infected mothers (n=7, n=9, n: biological replicates). T cells were identified as CD45+CD3+ and either CD4+ or CD8+ cells. Data information: In data are presented as mean ± SEM. *P≤0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001 B, Mann-Whitney-U test).Percentage of EM T cells (CD44+, CD62L-) among CD8+ T cells; (D) peripheral blood (n=6, n=9), (E) uterus-draining lymph nodes (n=6, n=9) n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001 (E, Student's t-test Mann-Whitney-U test).I-L. Percentage of ovalbumin-specific CD8+ EM T cells; (I) peripheral blood (n=7, n=9), (J) uterus-draining lymph nodes (n=7, n=9), (K) spleen (n=8 each), (L) uterus (n=6, n=9); n: biological replicates. Data information: I-L, data are presented as mean ± SEM. *P≤0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001 L: Student's t-test I: Mann-Whitney-U test).
Figure 2C-E. Percentage of CD3+ T cells of total MMc; (C) fetal bone marrow (n=13, n=14), (D) fetal spleen (n=12, n=13), (E) fetal liver (n=13, n=14); n: biological replicates. Data information: In C-E data are presented as mean ± SEM. *P≤0.05 (C, D, E Student's t-testG-I. percentage of OVA-specific MMc stained with OVA tetramers among CD3+ microchimeric T cells; (G) bone marrow (n=13 each), (H) spleen (n=12 each), (I) liver (n=11, n=12); n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05 H: Student's t-test; G, I: Mann-Whitney-U).J. Serum anti-Lm IgG titer of mothers and fetuses at gd 18.5 after preconceptual infection of the mother with LmOVA; n=3 biological replicates, each averaged from n=2 technical replicates). Data information data are presented as mean ± SEM. *P≤0.05
Figure 3Percentages of innate immune cell subsets in the fetal spleen; (B) CD45+ leukocytes (n=20, n=11), n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05 (B Ordinary one-way ANOVAPercentages of innate immune cell subsets in the fetal spleen (C) CD45+CD11b+ myeloid cells (n=19, n=11), (D) CD45+CD11b+Gr-1- monocytes (n=19, n=11), (E) CD45+CD11b+Gr-1+ neutrophils (n=19, n=11) and (F) CD45+CD11b+F4/80+ macrophages (n=20, n=11); n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05 Ordinary one-way ANOVA; D-F, : Kruskal-Wallis test).G-K. Percentages of innate immune cell subsets in the fetal liver; (G) CD45+ leukocytes (n=20, n=10), (H) CD45+CD11b+ myeloid cells (n=20, n=10), (I) CD45+CD11b+Gr-1- monocytes (n=20, n=10), (J) CD45+CD11b+Gr-1+ neutrophils (n=20, n=10) and (K) CD45+CD11b+F4/80+ macrophages (n=20, n=10); n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05 G-I: Ordinary one-way ANOVA K, Kruskal-Wallis test).
Figure 4C. Neonatal body weight in mg (n=20-55). (C) day 14 after birth (7 days after infection); n: biological replicates. D, E. Percentages of OVA tetramer+ cells among CD3+ MMc (percentages of OVA tetramer+ among CD3+ MMc at gd 18.5 (Fig 2h, i) are included for direct comparison); (D) spleen (n=5-10), (E) liver (n=5-10); n: biological replicates. Data information: data are presented as mean ± SEM. *P≤0.05; **P≤0.01; ****P≤0.0001 Ordinary one-way ANOVA; : Kruskal-Wallis test).F-I. Percentage of (F) CD3+ T cells of CD45+ spleen cells (n=6-14), (G) OVA-specific cells of CD3+ T cells from spleen cells (n=6-14), (H) CD11b+ cells of CD3+ T cells from spleen cells (n=6-14), and (I) CD11c+ cells of CD3+ T cells from spleen cells (n=6-14); n: biological replicates. Data information: In data are presented as mean ± SEM. *P≤0.05; **P≤0.01; ****P≤0.0001 G: Ordinary one-way ANOVA , H, I: Kruskal-Wallis test).
Figure 2E-H Experimental glucose uptake flux (E), acetate flux (F), and growth rate (G) of E. coli grown on glucose (15 mM) with different concentrations of αMG (0-100 mM), at low (0 mM) and high (30 mM) acetate concentrations. These data were used to quantify the total carbon uptake flux as a function of the αMG concentration (H). Each data point represents an independent biological replicate, and lines represent the best polynomial fits.I, J Total carbon uptake (I) and growth rate (J) as a function of the glucose uptake flux, at low (0 mM) and high (30 mM) acetate concentrations. Each data point represents an independent biological replicate, and lines represent the best linear fits. K, L Relationships between the acetate flux and glucose uptake (K), and between the contribution of acetate to carbon uptake and glucose uptake (L). Each data point represents an independent biological replicate, and lines represent the best polynomial fits.
Figure 3C Experimental acetate flux of E. coli K-12 MG1655 grown on glucose (15 mM) at different acetate concentrations in the presence of αMG (0, 20, 40 or 100 mM). Each data point represents an independent biological replicate, and the lines represent best fits using a logistic function.D Experimental acetate concentration threshold at which the acetate flux reverses as a function of the αMG concentration, determined from the best fits shown in panel (C).