EMBO/SourceData huggingface dataset:
Figure 4B Experimental growth rate of E. coli K-12 MG1655 grown on glucose (15 mM) as a function of the acetate concentration at high (0 mM αMG, light blue, low (100 mM αMG, dark blue) glycolytic flux. Each data point represents an independent biological replicate and the lines represent best polynomial fits.C Relative change in the growth rate of E. coli K-12 BW25113 wild-type in the presence of 10 mM acetate at high (0 mM αMG) and low (100 mM αMG) glycolytic flux. Mean values ± standard deviations (error bars) were estimated from n independent biological replicates, as indicated on the figure.D Relative change in the growth rates of E. coli K-12 BW25113 Δpgi and ΔpfkA strains in the presence of 10 mM acetate. Mean values ± standard deviations (error bars) were estimated from three independent biological replicates.
Figure 6Relative change in the growth rates of wild-type, Δpta, and Δacs E. coli K-12 BW25113 strains in the presence of 10 mM acetate at low glycolytic flux (100 mM αMG). Mean values and standard deviations (error bars) were estimated from n independent biological replicates, as indicated on the figure.
Figure 7A,B Impact of the presence of 10 mM acetate on the growth rate of wild-type, Δpta, and Δacs E. coli K-12 BW25113 strains grown on (A) 30 mM glycerol and (B) 15 mM galactose. Mean values and standard deviations (error bars) were estimated from three independent biological replicates.
Figure 1d/e: Density gradient centrifugation analysis of virus-containing supernatant. Forty milliliters of culture supernatant was harvested at 72 hours post ZIKV infection (MOI=0.01) and concentrated to 1 ml, followed by gradient centrifugation as described in the methods. Fractions were analyzed by Western blot with the indicated antibodies (d). The ZIKV titer of each fraction was measured by plaque assay, and the viral genome was determined via RT-PCR (e). Data represent the means from three biological replicates; error bars represent standard deviations from the means. Dashed line represents infectivity detection limit.f/g: Density gradient centrifugation analysis of UC-EVs. UC-EVs (0.1 ml) were loaded onto the top of a density gradient in a centrifugation tube as described in the methods. The fractions were obtained from the top of gradient and analyzed by Western blot with the indicated antibodies (f), the ZIKV titer of each fraction was measured by plaque assay, and the viral genome was quantified via RT-PCR (g). Data represent the means from three biological replicates; error bars represent standard deviations from the means. Dashed line represents infectivity detection limit.
Figure 2b: Western blots of ZIKV-infected cell-derived UC-EVs or IC-EVs. Samples during purification were subjected to western blot analysis with the indicated antibodies against cellular proteins. Total lysate of ZIKV-infected cells was served as positive control.d: Density gradient centrifugation analysis of IC-EVs. ZIKV-infected cell-derived UC-EVs (0.1 ml) were subjected to density gradient analysis similarly. Fractions were obtained from the top of gradient and analyzed by Western blot with the indicated antibodies.
Figure 3Detection of infectivity(d e) of UC-EVs pre- and post CD9-capture or E-capture. UC-EV was subjected to CD9-capture and E-capture respectively. After 1hour incubation, ZIKV infectivity of pre-incubation and post-incubation samples was measured.f: The ZIKV infectious titer of each fraction was measured by plaque assay, and the number of viral genomes was determined via RT-PCR. Data represent the means from three biological replicates; error bars represent standard deviations from the means. Dashed line represents infectivity detection limit.
Figure 4d: Western blots of ZIKV-infected cell-derived UC-EVs or IC-EVs using indicated antibodies against viral proteins and EV proteins. Total lysate of ZIKV-infected cells was the positive control.
Figure 5a: ZIKV E and CD9 distribution in a density gradient. After density gradient centrifugation, all fractions were analyzed by Western blotting with anti-ZIKV E and anti-CD9 antibodies. CD9 blots was reused from Figure 2d.d: Quantification of the number of ZIKV E proteins on the EV surface. The percentage of 10nm gold sphere-labeled IC-EVs was counted based on the immuno-EM images of IC-EVs. N indicates the total number of counted IC-EVs.
Figure 6c: Detection of ADE-induced ZIKV infection in the presence of the ZIKV-specific neutralizing antibody 2B10 and IC-EVs derived from ZIKV-infected cells by ADE assay. K562 cells were incubated with a mixture of EV-free ZIKV, neutralizing antibody, and IC-EVs. Forty-eight hours post incubation; cells were stained for ZIKV E and analyzed by FACS. Data represent the means from three biological replicates; error bars represent standard deviations from the means. Differences between treatments were analyzed using the two-tailed, unpaired Student t test available in the GraphPad Prism 5 software package. P values of less than 0.05 (indicated by asterisks) were considered statistically significant. The following categories were used: ***, P 0.001; **, P 0.01; *, P 0.05.
Figure 7c/d: IC-EVs affected the ADE of DENV2 (c) or ZIKV (d) mediated by DENV3 patient serum. Diluted sera were mixed with EV-free viruses (DENV2 or ZIKV) and IC-EVs derived from DENV2- or ZIKV-infected cells and then incubated with K562 cells as described in the methods. At 48 hours post incubation, cells were stained with an antibody against flavivirus E and analyzed by FACS. Data represent the means from three biological replicates; error bars represent standard deviations from the means. Differences between treatments were analyzed using the two-tailed, unpaired Student t test available in the GraphPad Prism 5 software package. P values of less than 0.05 (indicated by asterisks) were considered statistically significant. The following categories were used: ***, P 0.001; **, P 0.01; *, P 0.05.
Figure 1(B, Performance in the multi-trail inverted screen test is impaired in (B) SUR2-STOP1149 (WT n = 9 [3 male; 6 female]; STOP n = 9 [3 male; 6 female]; p = 0.011 according to Mann Whitney U test) Data information: Bars show mean (± SEM) with individual biological replicates shown as circles/squares. * denotes p value < α = 0.05 for all panels.C) Performance in the multi-trail inverted screen test is impaired in (C) SUR2-STOP475 mice (WT n = 9 [4 male; 5 female]; STOP n = 15 [5 male; 10 female]; p = 2.9x10-4 according to Mann Whitney U test). Data information: Bars show mean (± SEM) with individual biological replicates shown as circles/squares. * denotes p value < α = 0.05 for all panels.(D)Treadmill exercise tolerance is also reduced in SUR2-STOP475 mice (WT n = 4 [2 male; 2 female]; STOP n = 8 [4 male; 4 female]; p = 0.008 according to Mann Whitney U test). Treadmill workload tolerated was also reduced (WT 18.1 ± 2.2 vs STOP475 9.2 ± 0.9 J; p = 0.008 from Man Whitney U test), mean mouse body weight was WT 22.1 ± 0.9 vs STOP475 21.9 ± 0.6 g. Data information: Bars show mean (± SEM) with individual biological replicates shown as circles/squares. * denotes p value < α = 0.05 for all panels.
Figure 2(A) Example traces of inside-out patch clamp recordings of acutely isolated FDB myofibers of WT (black, top left), SUR2-STOP475 (purple, top right), and skeletal muscle dominant negative (SkM-DN; magenta, bottom left) mice. Patches were voltage clamped at -50 mV and excised at time-points indicated by arrows. ATP (5 mM) was administered to the cytoplasmic face of membrane patches as indicated by horizontal bars. Dashed lines denote the zero KATP current level.(B) Maximal ATP-sensitive current levels from excised patches. Bars show mean (± SEM) with individual recordings from each myofiber shown as dots/squares (cells from ≥ 3 mice; WT n = 10 recordings; STOP n = 8; SkM-DN n = 11; Dunn's test following Kruskal-Wallis omnibus test p = 1.0 x 10-4 for WT vs STOP and 0.014 for WT vs SkM-DN ; * denotes p value < adjusted α = 0.0167).(D) Distance travelled in treadmill exercise tolerance tests (all male; WT n = 6; AAA+ n = 8; Myf5-cre n = 8; SkM-DN n = 7; Dunn's test following Kruskal-Wallis omnibus test p = 0.005 for WT vs SkM-DN, 0.037 for AAA+ vs SkM-DN, and 0.014 for Myf5-cre vs SkM-DN; * denotes p value < adjusted α = 0.0083). Treadmill workload tolerated was also reduced (WT 23.6 ± 2.3 vs SkMDN 10.1 ± 1.8 J; p = 0.0017 from Dunn's test), mean mouse body weight was WT 28.9 ± 1.6, AAA+ 26.6 ± 0.9, Myf5-cre 25.3 ± 0.8, SkMDN 24.2 ± 0.8 g; no significant differences in body weights were observed, p from Dunn's test following Kruskal-Wallis omnibus test > adjusted α = 0.0083 for all comparisons. Bars show mean (± SEM) with individual mouse performance shown as dots/diamonds/triangles/squares.
Figure 3(A) Left - example HE stained tibialis anterior (TA) muscles from WT and SUR2-STOP475 mice. Black arrows identify peripheral nuclei and yellow arrows indicate centrally nucleated fibers. Right - quantification of centrally nucleated fibers. Bars show mean (± SEM) with individual biological replicates shown as dots/squares (WT n = 5; STOP n = 4; p = 0.016 according to Mann Whitney U test) (B) Left - example HE stained TA muscles from control and SkM-DN mice. Black arrows identify peripheral nuclei and yellow arrows indicate centrally nucleated fibers. Right - quantification of centrally nucleated fibers. Bars show mean (± SEM) with individual biological replicates shown as dots/squares (Control n = 7 [comprising 2 x WT, 3 x Myf5+-cre, and 2 x AAA+ littermate controls]; SkM-DN n = 5; p = 0.002 according to Mann Whitney U test). Data information: * denotes p value < α = 0.05 for all panels. Scale bars show 50 µm in each panel.(C) Example of HE stained WT TA muscle (left) and two examples of HE stained SkM-DN TA muscle alongside immunolabelling showing presence of CD68+/CD206+ (macrophage marker) cells indicated by green arrows.(D) Left - Picrosirius red stained TA muscle from WT and SkM-DN mice. Right - quantification of red stained area (WT n = 4; SkM-DN n = 4; p = 0.028 according to Mann Whitney U test). Bars show mean (± SEM) with individual measurements shown as dots/squares. (E) Left - TA muscle for WT and SkM-DN mice showing Pax7 positive immunolabelled satellite cells (red arrows). Right - quantification of Pax7+ cells (WT n = 4; SkM-DN n = 4; p = 0.028 according to Mann Whitney U test). Bars show mean (± SEM) with individual measurements shown as dots/squares. Data information: * denotes p value < α = 0.05 for all panels. Scale bars show 50 µm in each panel.
Figure 4(A) Example traces showing force generation in isometric contractility studies of isolated EDL muscle from WT (left, black) and SUR2-STOP475 (right, purple) mice. Muscle was stimulated every 2 s. Scale bars show 20 mN (y-axis) and 2 s (x-axis). (B) Enlarged representations of the 1st (at 0 seconds), 10th (20 s) and 30th (60 s) contractions, identified in concatenated traces in (A) by arrows. Scale bars show 20 mN (y-axis) and 0.4 s (x-axis).
Figure 5(A) Survival of WT and SUR2-STOP475 mice administered with verapamil in drinking water (n = 7 [4 male, 3 female] for WT administered 0.9 g/l verapamil; n = 14 [8 male, 6 female] for SUR2-STOP475 administered 0.9 g/l verapamil; n = 6 [3 male, 3 female] for SUR2-STOP475 administered 0.3 g/l verapamil). Verapamil induced deaths were observed in both male and female mice (7 of 8 male and 4 of 6 female SUR2-STOP475 mice died within 28 days of 0.9 g/l verapamil). (B) Survival of WT and SUR2-STOP1149 mice administered with verapamil in drinking water (n = 10 [3 male, 7 female] for SUR2-STOP1149 mice administered 0.9 g/l verapamil).(D) Heart rates calculated from ECG recordings from WT and SUR2-STOP mice administered with no verapamil (0 Verap) or 0.9 g/l verapamil in drinking water.(F) Cumulative durations of inversion in multi-trial inverted screen test for WT mice (black circles) and SkM-DN mice without (magenta squares) or with 0.9 g/l verapamil (pink triangles) administration. Data from WT and SkM-DN without verapamil Bars show mean ± SEM (WT 0 verapamil n = 9 [6 male; 3 female]; SkM-DN 0 verapamil n = 16 [12 male; 4 female]; SkM-DN with 0.9 g/l verapamil n = 6 [4 male; 2 female]; Dunn's test p = 0.006 for WT 0 verapamil vs SkM-DN 0 verapamil and 0.027 for WT 0 verapamil vs SkM-DN 0.9 g/l verapamil following Kruskal-Wallis omnibus test; * denotes p value < adjusted α = 0.017).
Figure 6(A) Left - Average performance in the inverted screen test of mice homozygous for the ncDHPR allele alone (ncDHPR) or homozygous for both the ncDHPR and SUR2-STOP475 alleles (STOP475/ncDHPR). Right - Summary of inverted screen test data. Data shown as mean ± SEM (WT n = 9 [4 male; 5 female]; ncDHPR n = 15 [male 6; female 9]; STOP475 n = 15 [5 male; 10 female]; STOP475/ncDHPR n = 12 [4 male; 12 female]; Dunn's test p = 4.0 x 10-4 for WT vs STOP475, 6.9 x 10-4 for WT vs STOP475/ncDHPR, 0.001 for ncDHPR vs STOP475/ncDHPR verapamil, following Kruskal-Wallis omnibus test). * denotes p value < adjusted α = 0.0083.(B) Left - Example of HE stained tibialis anterior muscle from STOP475/ncDHPR mouse, black arrow shows peripheral nuclei and yellow arrows show central nuclei. Right - Quantification of centrally nucleated fibers, data from WT and SUR2-STOP475 mice (WT n = 5; STOP475 n = 4; STOP475/ncDHPR n = 8; Dunn's test p = 0.007 for WT vs STOP475 and 0.011 for WT vs STOP475/ncDHPR, following Kruskal-Wallis test, * denotes p value < adjusted α = 0.067). Scale bar shows 50 µm. Bars show mean (± SEM) with individual measurements shown as dots/squares/triangles.(F) Summary of unstimulated force (normalised by physiological cross-sectional area) at 60 s for each experiment. Bars show mean ± SEM with individual biological replicates shown as dots/squares/triangles (WT without glibenclamide n = 6 [3 male; 3 female]; WT with glibenclamide n = 10 [5 male; 5 female]; ncDHPR without glibenclamide n = 6 [3 male; 3 female]; ncDHPR with glibenclamide n = 6 [3 male; 3 female]; Dunn's test p = 7.0 x 10-4 for WT without glibenclamide vs WT with glibenclamide and 0.002 for ncDHPR without glibenclamide vs ncDHPR with glibenclamide, following Kruskal-Wallis omnibus test; * denotes p value < adjusted α = 0.0083).
Figure 1Serum cytokine levels from control-treated (black) and anti-HER2/CD3 TDB-treated (red) mice, 6 h and 24 h post treatment (n = 9 mice/treatment).G, H. Representative images of tumor (G) or liver (H) sections from the Founder5 model stained with CD3 (red), Endomucin (white), and DAPI (blue). Right panels: quantification of CD3+ cell counts in Founder5 whole tumor (G) or liver (H) sections normalized to tissue areas. All samples were analyzed 24 h post treatment, n = 8/9 mice/treatment.Tumor (I) tissue cytokine levels normalized to total protein concentration from control-treated (black) and anti-HER2/CD3 TDB-treated (red) mice, 24 h post treatment (n = 8 mice/treatment).
Figure 2B, C. Flow cytometry analysis of T cell phenotype and T cell subsets for CD8+ T cells (B) and CD4+ T cells (C) in dissociated tumors normalized to tumor weights (upper panels) or dissociated livers normalized to liver weights (lower panels), 24 h post treatment. (n = 8 mice/treatment).
Figure 3Cytokine concentrations in conditioned medium of HUVECs cultured without treatment (blue diamond, treatment b), or with 600 pg/ml a (pink circle, treatment c), or with control PBMC supernatant (orange triangle, treatment d), or with TDB-conditioned PBMC supernatant (purple square, treatment e). All done in n = 6 replicates with 6 hours incubation. Endothelial cell culture medium alone (black circle, treatment a, n = 4) was plotted as reference.Gating strategy for flow cytometry analysis of surface expression of ICAM-1 and VCAM-1 on HUVECs. Singlets were gated based on forward scatter area (FSC-A) versus forward scatter height (FSC-H). Activated endothelial cells were gated as ICAM-1+ or VCAM-1+ endothelial cells from viable cells.   Quantification of percentage of ICAM-1+ (top panel) and VCAM-1+ (bottom panel) endothelial cells with the same treatments as defined in panel B. All done in triplicates.
Figure 4A, B. Representative images of liver (A) or tumor (B) sections from the Founder5 tumor-bearing mice stained with ICAM‑1 (red), VCAM-1 (white), and Endomucin (green) 24 hours post treatment. Right panels are enlarged views of the boxed areas showing only ICAM-1 and VCAM-1. Scale bar = 100 μm. Below: quantification of total ICAM-1+ and VCAM-1+ areas as well as vascular (vasc.) ICAM‑1+ (ICAM-1+ Endomucin+) and vascular VCAM-1+ (VCAM‑1+ Endomucin+) areas normalized to tissue areas. n = 9 mice/treatment.E, F. Densities of total blood vessels (left panel) and mature blood vessels (right panel) in liver (E) and tumor (F) sections from the Founder5 tumor-bearing mice normalized to tissue areas. n = 9 mice/treatment.G, H. Left panels: representative images of liver (G) or tumor (H) sections from the Founder5 model stained with ICAM‑1 (white), pericyte markers (pink), and Endomucin (green) (top panel) or stained with VCAM‑1 (white), pericyte markers (pink), and Endomucin (green) (bottom panel) 24 hours post treatment. Scale bar = 100 μm. Right panels: linear regression analysis comparing adhesion molecule (ICAM-1 and VCAM-1) expression (x-axis) and vessel maturity (y-axis). The coefficients of determination (R) are indicated. n = 9 mice/treatment.
Figure 6C, D. Representative images of liver (C) or tumor (D) sections from the Founder5 model stained with the following combinations of markers: (left panels) CD9 (green) and ICAM-1 (red); (right panels) CD9 (green) and CD31 (white); 24 hours post treatment. In (D) the 3rd right panels are enlarged views of the boxed areas showing CD9+ cells inside the lumen of blood vessels but not in the endothelium; orthogonal views (z) of selected planes are shown on the far right.Quantitative RT-qPCR analysis of relative expression of Cd9 normalized to Gapdh in HUVEC after stimulation with either control (blue diamond, n = 9) or PBMC TDB-conditioned supernatant (lilac square, n = 3) for 6 h and 24 h.Quantification of T cell adhesion to a monolayer of HUVECs transfected with either negative control siRNA (black) or CD9-specifc siRNA (green) under shear stress-like conditions (100 rpm) treated with the indicated conditions. All done in biological duplicates with four technical replicates per biological replicate. Representative data of one experiment shown. FOV: field of view.
Figure 1(B) Dose-dependent inhibition of nirmatrelvir (left panel) and GC376 (right panel) on SARS-CoV-2 Mpro. Prior to adding the substrate for the biochemical reaction, the protease was preincubated for 30 minutes at 37°C with the indicated concentrations of the compound.(C) Dose-dependent antiviral activity of nirmatrelvir in HEK293T-hACE2 cells infected with SARS-CoV-2 D614G (purple symbols), B.1.617.2 (green symbols) or B.1.1.529 (blue symbols). Antiviral activity was determined as percent inhibition of the virus-induced cytopathic effect.(F) Quantification of SARS-CoV-2 RNA (left panel) and viral titers (right panel) in the lung 3 days after infection. RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. Viral titers were determined by median tissue culture infectious dose (TCID50).(G) Quantification of SARS-CoV-2 RNA in the nasal turbinates 3 days after infection. RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line.
Figure 2Quantification of anti-S1 RBD IgG levels by ELISA in the plasma of the indicated mice (C) 7, 14 and 21 days p.i. Neutralization dose 50 (ND50) against SARS-CoV-2 B.1.1.529 pseudovirus in the plasma of the indicated mice (D)Quantification of anti-S1 RBD IgG levels by ELISA in the plasma of the indicated mice (E) 4 days post re-challenge. Neutralization dose 50 (ND50) against SARS-CoV-2 B.1.1.529 pseudovirus in the plasma of the indicated mice (F) 4 days post re-challenge.(I-N) Representative flow cytometry plots of (I) CD8+ T cells or (L) CD4+ T cells expressing IFN-γ and TNF-α in the lungs of the indicated mice 4 days post re-challenge. Unstimulated cells are shown in the upper panels, whereas cells re-stimulated with a pool of SARS-CoV-2 peptides for 4 hours at 37°C are shown in the bottom panels. Plots were pre-gated as (I) live+/ B220-/CD19-/CD4-/CD8+ cells or (L) live+/B220-/CD19-/CD8-/CD4+. Frequency (J, M) and absolute number (K, N) of IFN-γ− and TNF-α−producing CD8+ T cells (J, K) or CD4+ T cells (M, N) in the lung of the indicated mice 4 days post re-challenge.
Figure 3(B, C) Quantification of anti-S1 RBD IgG levels by ELISA in the plasma of the indicated mice (B) 14, 21 days p.i. and (C) 4 days post re-challenge.(E) Quantification of anti-S1 RBD IgG levels by ELISA in the plasma of the indicated mice 7 days p.i..(F) Flow cytometry histogram (left panel) and geometric mean fluorescent intensity (gMFI) quantification (right panel) of CD44 expression by CD8+ T cells in the blood of the indicated mice 7 days p.i. (pre-gated on live+/ B220-/ CD19-/ CD4-/ CD8+ cells).(G, H) Frequency of (G) Granzyme-B− and (H) IFN-γ−producing CD8+ T cells in the blood of the indicated mice 7 days p.i.. Cells were stimulated in vitro with a pool of SARS-CoV-2 peptides for 4 hours at 37°C. Plots were pre-gated as above.(J) Quantification of anti-S1 RBD IgG levels by ELISA in the plasma of the indicated mice 7, 14 and 21 days p.i..
Figure 3CAC was induced in control (n = 7) and ColVIcre-Kif3aflx/flx (n = 7) female mice according to the timeline shown in Figure 2A. (E) Representative images of hematoxylin and eosin staining of paraffin-embedded colon sections. Areas with high-grade dysplasia are delimited by black lines.CAC was induced in control (n = 7) and ColVIcre-Kif3aflx/flx (n = 7) female mice according to the timeline shown in Figure 2A. (F,G,H,I) Box-and-whisker plots depicting area (F,G) and number (H,I) of low- and high-grade dysplasia per mouse.
Figure 4(D,E,F) At the end of the protocol described in (A) mice were sacrificed and colons were analyzed by histology for colitis severity including areas of irregular crypts (i.e., non-parallel crypts, variable crypt diameters, bifurcation and branched crypts; D,E) and crypt loss (i.e. mucosa devoid of crypts; D,F).(H) Elevated numbers of F4/80+ macrophages in colons of DSS-treated ColVIcre-Kif3aflx/flx mice. Representative images for F4/80 staining in areas with crypt loss are shown. At least 5 fields in the regions of crypt loss were analyzed from each colon of control (n=6) and ColVIcre-Kif3aflx/flx (n=6) mice. Mean cell numbers were scored as low (<500 cells/mm2) or high (>500 cells/mm2).(N) Decreased number of PC in inflamed tissue of patients with ulcerative colitis. Shown are the results of matched-pair analysis for PC expression per vimentin+ cells as arbitrary unit (AU, see material and methods). The lines link values obtained for regions of ulceration in the ileum with those of normal regions from the same patient, demonstrating the difference in each pair.
Figure 5Elevated numbers of IL-6-expressing F4/80+ macrophages in ColVIcre-Kif3aflx/flx mice treated with DSS as described in Figure 4A. At least 5 fields in the regions of crypt loss were analyzed. Scale bar represent 100 µm. *p<0,05 was calculated by chi-squared test.Elevated IL-6 expression in colonic epithelial cells and fibroblasts of PC-deficient mice upon DSS-treatment as described in Figure 4A. Representative images and corresponding quantification of vimentinpositive fibroblasts (in green) and vimentinnegative epithelial cells (identified by histological criteria) labeled for IL-6 (upper panel) and p-STAT3 (lower panel) in red. Nuclei were stained with DAPI (in blue). White arrow heads indicate IL-6 (upper panel) and nuclear staining for pSTAT3 (lower panel) for vimentinpositive fibroblasts, blue arrow heads the respective labeling for epithelial cells. Scale bars represent 100 µm. *p<0.05, ****p<10-4, n.s. (not significant) was calculated by chi-squared test on absolute numbers and is presented as % of cells positive for the indicated marker within vimentinpositive and epithelial cells, respectively.