Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Experimental Study on Danggui Shaoyao San Improving Renal Fibrosis by Promoting Autophagy

doi: 10.1155/2022/6761453

Figure Lengend Snippet: Selection of DSS treatment concentration. (a) NRK-52E cells were treated, respectively, with different drug concentrations of DSS (0, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4, and 12.8 mg/ml) for different time periods (24 h, 48 h, and 72 h) ( n = 3, per group). (b) NRK-52E cells were treated, respectively, with different drug concentrations of DSS (0, 0.2, 0.4, 0.8, 1.6, and 3.2 mg/ml) at 5 ng/ml TGF- β 1 for 48 h ( n = 5, per group). Then, NRK-52E cells were treated, respectively, with 5 ng/ml TGF- β 1 and different drug concentrations of DSS ( n = 3, per group). “+” means containing the drug, “--” means without the reagent, “0.4, 0.8, 1.6” means the used drug concentration of DSS (mg/ml). (c) Protein levels of LC3 I, LC3 II, fibronectin, and caspase-3 in cellular were determined by western blotting. (d) Quantification of protein levels. Results are expressed as mean SEM. # p < 0.05 versus control; ∗ p < 0.05 and ∗∗ p < 0.01 versus TGF- β 1 only.

Article Snippet: The membranes were probed overnight at 4°C with primary antibodies against fibronectin (Abcam, Cambridge, CB, UK), caspase-3(Abcam, Cambridge, CB, UK), Phospho-mTOR (CST, Danvers, MA, USA), mTOR (Abcam, Cambridge, CB, UK), Beclin 1(Abcam, Cambridge, CB, UK), LC3B (Abcam, Cambridge, CB, UK), p62(CST, Danvers, MA, USA), GAPDH (Proteintch, Rosemont, IL, USA), and β -actin (Proteintch, Rosemont, IL, USA).

Techniques: Selection, Concentration Assay, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Experimental Study on Danggui Shaoyao San Improving Renal Fibrosis by Promoting Autophagy

doi: 10.1155/2022/6761453

Figure Lengend Snippet: DSS reduced FN secretion and apoptosis of NRK-52E cells induced by TGF- β 1. NRK-52E cells were treated, respectively, with TGF- β 1(5 ng/ml), DSS (0.8 mg/ml) + TGF- β 1(5 ng/ml), and rapamycin (10 nM) + TGF- β 1(5 ng/ml) for 48 h ( n = 3, per group). (a) Protein levels of fibronectin and caspase-3 in cellular were determined by western blotting. (b) Quantification of protein levels. Results are expressed as mean SEM. # p < 0.05 versus control; ∗ p < 0.05 versus TGF- β 1 only.

Article Snippet: The membranes were probed overnight at 4°C with primary antibodies against fibronectin (Abcam, Cambridge, CB, UK), caspase-3(Abcam, Cambridge, CB, UK), Phospho-mTOR (CST, Danvers, MA, USA), mTOR (Abcam, Cambridge, CB, UK), Beclin 1(Abcam, Cambridge, CB, UK), LC3B (Abcam, Cambridge, CB, UK), p62(CST, Danvers, MA, USA), GAPDH (Proteintch, Rosemont, IL, USA), and β -actin (Proteintch, Rosemont, IL, USA).

Techniques: Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Trichostatin A Alleviates Renal Interstitial Fibrosis Through Modulation of the M2 Macrophage Subpopulation

doi: 10.3390/ijms21175966

Figure Lengend Snippet: Macrophage infiltration correlates with renal interstitial fibrosis following a unilateral ureteral obstruction (UUO) injury. ( A ) Representative images of the Masson trichrome staining showed that the extent of fibrosis (blue staining) progressively increased in day 14 UUO kidneys as compared to day 7 ones. Immunohistochemistry demonstrated that more α-smooth muscle actin (α-SMA)-positive myofibroblasts and F4/80-positive macrophages accumulated in day 14 UUO kidneys. Scale bar = 100 μm. ( B ) Quantification of the fibrosis extent, α-SMA-positive and F4/80-positive areas. ** p < 0.01 and *** p < 0.001 by the unpaired Student’s t -test; n = 5 for each group. ( C ) Western blot analysis of fibronectin and α-SMA expression in day 7 and day 14 obstructed kidneys. β-actin served as the loading control. ( D ) Quantification of the Western blot analyses. ** p < 0.01 and *** p < 0.001 by the unpaired Student’s t -test; n = 5 for each group.

Article Snippet: The membranes were then probed with primary antibodies against fibronectin (1:1000, Cat#15613-1-AP, Proteintech Group, Chicago, IL, USA), α-SMA (1:5000, Cat#14395-1-AP, Proteintech Group), iNOS (1:1000, Cat#ab3523, Abcam), Arg1 (1:1000, Cat#93668S, Cell Signaling Technology, Boston, MA, USA), CLEC7A (1:500, Cat#TA322197, OriGene Technologies), SLAM (1:1000, Cat#ab156288, Abcam), TNF-α (1:1000, Cat#ab66579, Abcam) and β-actin (1:5000, Cat#60008, Proteintech Group) at 4 °C overnight.

Techniques: Staining, Immunohistochemistry, Western Blot, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Trichostatin A Alleviates Renal Interstitial Fibrosis Through Modulation of the M2 Macrophage Subpopulation

doi: 10.3390/ijms21175966

Figure Lengend Snippet: Trichostatin A (TSA) ameliorates renal inflammation and fibrosis in unilateral ureteral obstruction (UUO). ( A ) Representative images of Masson trichrome staining of obstructed kidneys at 7 days and 14 days following UUO in the vehicle and TSA groups. The TSA treatment markedly reduced the extent of interstitial fibrosis (blue area). Immunohistochemical staining for α-smooth muscle actin (α-SMA, myofibroblast marker) and F4/80 (pan-macrophage marker) in day 7 and day 14 obstructed kidneys. Scale bar = 100 μm. ( B ) Quantification of the fibrosis extent, α-SMA-positive and F4/80-positive areas. * p < 0.05, ** p < 0.01 and *** p < 0.001 by the unpaired Student’s t -test; n = 5 for each group. ( C ) Western blots of fibronectin and α-SMA in day 7 and day 14 UUO kidneys treated with the vehicle or TSA. β-actin served as the loading control. ( D ) Quantification of fibronectin and α-SMA expression levels. ** p < 0.01 by the unpaired Student’s t -test; n = 5 for each group.

Article Snippet: The membranes were then probed with primary antibodies against fibronectin (1:1000, Cat#15613-1-AP, Proteintech Group, Chicago, IL, USA), α-SMA (1:5000, Cat#14395-1-AP, Proteintech Group), iNOS (1:1000, Cat#ab3523, Abcam), Arg1 (1:1000, Cat#93668S, Cell Signaling Technology, Boston, MA, USA), CLEC7A (1:500, Cat#TA322197, OriGene Technologies), SLAM (1:1000, Cat#ab156288, Abcam), TNF-α (1:1000, Cat#ab66579, Abcam) and β-actin (1:5000, Cat#60008, Proteintech Group) at 4 °C overnight.

Techniques: Staining, Immunohistochemical staining, Marker, Western Blot, Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Trichostatin A Alleviates Renal Interstitial Fibrosis Through Modulation of the M2 Macrophage Subpopulation

doi: 10.3390/ijms21175966

Figure Lengend Snippet: The effect of trichostatin A (TSA) on proinflammatory and profibrotic phenotypes of M2a and M2c macrophages. ( A ) Western blots of C-type lectin domain family 7 member A (CLEC7A), proinflammatory (tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS)) and profibrotic (α-smooth muscle actin (α-SMA) and fibronectin) expressions in J774A.1 macrophages treated with the indicated conditions for 24 h. Concentrations of interleukin (IL)-4 and IL-13 were both 20 ng/mL. β-actin served as the loading control. ( B ) Quantification for the levels of indicated proteins. * p < 0.05 and ** p < 0.01 by ANOVA, followed by Tukey’s post hoc multiple comparison test; n = 3 for each group. ( C ) Western blots of SLAM, proinflammatory (TNF-α and iNOS) and profibrotic (α-SMA and fibronectin) expressions in J774A.1 macrophages treated with the indicated conditions for 24 h. Concentrations of IL-10 and transforming growth factor (TGF)-β1 were both 20 ng/mL. β-actin served as the loading control. ( D ) Quantification for the levels of indicated proteins. * p < 0.05 and ** p < 0.01 by ANOVA, followed by Tukey’s post hoc multiple comparison test; n = 3 for each group.

Article Snippet: The membranes were then probed with primary antibodies against fibronectin (1:1000, Cat#15613-1-AP, Proteintech Group, Chicago, IL, USA), α-SMA (1:5000, Cat#14395-1-AP, Proteintech Group), iNOS (1:1000, Cat#ab3523, Abcam), Arg1 (1:1000, Cat#93668S, Cell Signaling Technology, Boston, MA, USA), CLEC7A (1:500, Cat#TA322197, OriGene Technologies), SLAM (1:1000, Cat#ab156288, Abcam), TNF-α (1:1000, Cat#ab66579, Abcam) and β-actin (1:5000, Cat#60008, Proteintech Group) at 4 °C overnight.

Techniques: Western Blot, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Trichostatin A Alleviates Renal Interstitial Fibrosis Through Modulation of the M2 Macrophage Subpopulation

doi: 10.3390/ijms21175966

Figure Lengend Snippet: The effect of trichostatin A (TSA) on the activation of renal myofibroblasts. ( A ) Western blots of fibronectin and α-smooth muscle actin (a-SMA) expressions in transforming growth factor-β1 (TGF- β1, 20 ng/mL)-stimulated renal tubular epithelial NRK-52E cells at the indicated time. β-actin served as the loading control. ( B ) Quantification for the levels of indicated proteins. * p < 0.05, ** p < 0.01 and *** p < 0.001 by ANOVA, followed by Tukey’s post hoc multiple comparison test. ns, nonsignificance. n = 3 for each group. ( C,D ) Immunofluorescent staining of fibronectin and α-SMA in the TGF-β1-treated NRK-52E cells at the indicated time. Scale bar = 50 μm. * p < 0.05, ** p < 0.01 and *** p < 0.001 by ANOVA, followed by Tukey’s post hoc multiple comparison test. ns, nonsignificance. n = 3 for each group.

Article Snippet: The membranes were then probed with primary antibodies against fibronectin (1:1000, Cat#15613-1-AP, Proteintech Group, Chicago, IL, USA), α-SMA (1:5000, Cat#14395-1-AP, Proteintech Group), iNOS (1:1000, Cat#ab3523, Abcam), Arg1 (1:1000, Cat#93668S, Cell Signaling Technology, Boston, MA, USA), CLEC7A (1:500, Cat#TA322197, OriGene Technologies), SLAM (1:1000, Cat#ab156288, Abcam), TNF-α (1:1000, Cat#ab66579, Abcam) and β-actin (1:5000, Cat#60008, Proteintech Group) at 4 °C overnight.

Techniques: Activation Assay, Western Blot, Control, Comparison, Staining

Journal: British Journal of Cancer

Article Title: Integrin α 3 is overexpressed in glioma stem-like cells and promotes invasion

doi: 10.1038/bjc.2013.218

Figure Lengend Snippet: Neurospheres formed by glioblastoma surgical specimens. ( A ) Neurospheres formed by glioblastoma surgical specimens (GBM61 and GBM69) in serum-free culture media containing 20 ng ml −1 of EGF and bFGF were immunostained with antibodies against glioma stem-like cell marker (CD133 and nestin). Note that the CD133 and nestin were immunostained in the cells. ( B ) Neurospheres from glioblastoma surgical specimens (GBM61 and GBM69) were cultured on plates coated with fibronectin (10 μ g ml −1 ) or laminin (10 μ g ml −1 ) in serum-free culture media containing 20 ng ml −1 of EGF and bFGF for 72 h. (original magnification × 100).

Article Snippet: Chemotactic migration on fibronectin and laminin and invasion into Matrigel (Becton Dickinson, Franklin Lakes, NJ) of cells was measured in a modified Boyden chamber (Neuroprobe, Gaithersburg, MD, USA) as described previously ( Yoshida et al , 2010 ).

Techniques: Marker, Cell Culture

Journal: British Journal of Cancer

Article Title: Integrin α 3 is overexpressed in glioma stem-like cells and promotes invasion

doi: 10.1038/bjc.2013.218

Figure Lengend Snippet: Analysis of cell migration of glioma cell lines with knock down of integrin α 3. ( A ) The relative mRNA expression levels of integrin α 3 (ITGA3) were calculated by quantitative RT-PCR. The value in U87 cells was taken as 1. Extracts of U87, U251, T98G, and SNB19 cells were immunoblotted with antibodies against integrin α 3 (ITGA3) or β -actin. ( B ) Extracts of U87, SNB19, or U251 cells treated by siRNA for integrin α 3 (siITGA3-1 and siITGA-2) or control luciferase (control) were immunoblotted with antibodies against integrin α 3 (ITGA3) or β -actin. ( C ) Cells treated by siRNA were plated on membranes coated with fibronectin (upper panel) or laminin (lower panel) in a chemotaxis chamber. Cells migrated to the underside of membrane for 6–8 h, and the migrated cells were stained with Diff-Quick and the absorbance measured at 490 nm as described in the Materials and Methods section. The mean absorbance (490 nm) value from mock cells in each cell line is shown as 1. Bar: mean±s.e. ( n =6). Double asterisks indicate a significant difference from the control at a P -value of <0.01.

Article Snippet: Chemotactic migration on fibronectin and laminin and invasion into Matrigel (Becton Dickinson, Franklin Lakes, NJ) of cells was measured in a modified Boyden chamber (Neuroprobe, Gaithersburg, MD, USA) as described previously ( Yoshida et al , 2010 ).

Techniques: Migration, Knockdown, Expressing, Quantitative RT-PCR, Control, Luciferase, Chemotaxis Assay, Membrane, Staining, Diff-Quik

Journal: British Journal of Cancer

Article Title: Integrin α 3 is overexpressed in glioma stem-like cells and promotes invasion

doi: 10.1038/bjc.2013.218

Figure Lengend Snippet: Analysis of cell migration and invasion of glioma cell lines upon manipulating the gene expression of integrin α 3. ( A ) Extracts of U87, SNB19, or U251 cells transiently transfected with integrin α 3 (ITGA3) or empty plasmid vector (Mock) were immunoblotted with antibodies against ITGA3 or β -actin. ( B ) Transfected cells were plated on membranes coated with fibronectin (upper panel) or laminin (lower panel) in a chemotaxis chamber. Bar: mean±s.e. ( n =6). Asterisk indicates a significant difference from the control at a P -value of <0.05. ( C ) U87 cells transfected with integrin α 3 (ITGA3) or empty plasmid vector (Mock) (upper panel) and treated by siRNA for integrin α 3 (siITGA3-1 and siITGA3-2) or luciferase (lower panel) were plated on membranes coated with Matrigel in a chemotaxis chamber. Cells were allowed to migrate to the underside surface of membranes for 24 h; invading cells were stained with Diff-Quick and the absorbance was measured at 490 nm, as described in Materials and Methods. The mean absorbance (490 nm) value from mock or control cells in each figure is shown as 1. Bar: mean±s.e. ( n =6). Single and double asterisks indicate significant differences between mock and control at P -values of <0.05 and <0.01, respectively.

Article Snippet: Chemotactic migration on fibronectin and laminin and invasion into Matrigel (Becton Dickinson, Franklin Lakes, NJ) of cells was measured in a modified Boyden chamber (Neuroprobe, Gaithersburg, MD, USA) as described previously ( Yoshida et al , 2010 ).

Techniques: Migration, Gene Expression, Transfection, Plasmid Preparation, Chemotaxis Assay, Control, Luciferase, Staining, Diff-Quik

Journal: British Journal of Cancer

Article Title: Integrin α 3 is overexpressed in glioma stem-like cells and promotes invasion

doi: 10.1038/bjc.2013.218

Figure Lengend Snippet: Effect of integrin α 3 on ERK1/2 and Rac activation. ( A ) U251 cells transiently transfected with integrin α 3 (ITGA3) or empty plasmid vector (Mock) and U87 cells treated with siRNA against integrin α 3 (siITGA3-1, siITGA3-2) or control luciferase (Control) were grown for 24 or 48 h. After starvation for 12 h, the cells were seeded on plates coated with fibronectin (10 μ g ml −1 ) and were incubated for 30 min. Extracts of the cells were immunoblotted with antibodies against phosphorylated ERK1/2 (p-ERK1/2), total ERK1/2 (ERK1/2), integrin α 3 (ITGA3), or β -actin. ( B ) U251 cells transiently transfected with integrin α 3 (ITGA3) or empty plasmid vector (Mock) were plated on membranes coated with Matrigel in a chemotaxis chamber. Cells were allowed to migrate for 24 h in the absence or presence of 20 μ ℳ PD98059 to the underside of the membranes, and the invading cells were measured. Mean cell counts from at least six fields in each of the four experiments are shown. Bars represent s.e. * P <0.05; ** P <0.01 vs Mock without PD98059.

Article Snippet: Chemotactic migration on fibronectin and laminin and invasion into Matrigel (Becton Dickinson, Franklin Lakes, NJ) of cells was measured in a modified Boyden chamber (Neuroprobe, Gaithersburg, MD, USA) as described previously ( Yoshida et al , 2010 ).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Control, Luciferase, Incubation, Chemotaxis Assay

Journal: Nature cell biology

Article Title: Two succeeding fibroblastic lineages drive dermal development and the transition from regeneration to scarring.

doi: 10.1038/s41556-018-0073-8

Figure Lengend Snippet: Fig. 2 | EPFs direct dermis development. a–e, Coronal sections of E9.5–E11.5 En1Cre;R26VT2/GK3 embryos. Arrows indicate single EPFs (b,d). Dashed line indicates the ectoderm (ec) border (b). Dotted lines delimitate EPFs clones (c–e). f, E12.5 En1Cre;R26VT2/GK3 embryo showing that EPFs arc across the back- skin. f’ and f” indicate high magnification from boxes in f. Arrows indicate migrating EPFs. The cross indicates the embryonic axes. Ant, anterior; D, dorsal; Pos, posterior; V, ventral. g–i, Immunofluorescence for fibronectin (g), collagen I (h) or collagen III (i) in coronal sections of E9.5 or E11.5 En1Cre;R26VT2/GK3 embryos. Dotted lines delimitate EPFs. j, Orthogonal view from time-lapse images (0 h and 24 h) from the anterior region of an E12.5 En1Cre;R26mTmG embryo. Dotted lines in the x–y frame delimitate the EPF-induced ENF gap. The green channel (EPFs) was omitted in x–y frames for clarity. Dotted lines from x–z and y–z delimitate EPFs. Dashed lines mark the dorsal surface. Arrows indicate the movement direction of EPFs. Scale bars, 50 µm (a–d), 500 µm (e,f), 200 µm (f',f'',g–i) and 100 µm (j). Images in a–j are representative of three experiments.

Article Snippet: Antibodies used: Ki67 (Abcam), Cas3 (Cell Signaling), collagen I (Rockland), fibronectin (Abcam), FSP1 (Abcam), collagen III (Abcam), collagen VI (Abcam), CD31 (Novus Biologicals), α -SMA (Abcam), CD26 (Abcam), TNC (Abcam), Dlk1 (Abcam), Sca1 (Biolegend), FABP4 (Abcam) and AF647 secondary antibody (Molecular Probes).

Techniques: Clone Assay, Immunofluorescence

Journal: Nature cell biology

Article Title: Two succeeding fibroblastic lineages drive dermal development and the transition from regeneration to scarring.

doi: 10.1038/s41556-018-0073-8

Figure Lengend Snippet: Fig. 4 | Dermal lattice is actively changing during development. a,b, Fractal dimension (FD) values (a) and lacunarity (L) values (b) derived from images of EPFs from different regions and developmental stages. Mean ± s.e.m. Repeated measures ANOVA, Newman–Keuls test, n = 9 optical fields of anterior or posterior regions at E11.5, E12.5, E14.5 and E16.5. *P < 0.05, **P < 0.01 and ***P < 0.001. c, 3D reconstructions of posterior and anterior En1Cre;R26VT2/GK3 EPFs. d, Distances (µm) between fibronectin fibres calculated from linear profiles of three confocal images of fibronectin-immunolabelled sections (Supplementary Fig. 6; n = 184 (E11.5 anterior), 138 (E11.5 posterior), 135 (E12.5 anterior), 155 (E12.5 posterior), 98 (E14.5 anterior), 105 (E14.5 posterior), 91 (E16.5 anterior), 128 (E16.5 posterior) measurements), mean ± s.e.m., ANOVA, Holm–Sidak test, *P < 0.05. e,f, FD values (e) and L values (f) derived from fibronectin stainings from different regions and developmental stages. Mean ± s.e.m. Repeated measures ANOVA, Newman–Keuls test, n = 3 optical fields of anterior or posterior regions at E11.5, E12.5, E14.5 and E16.5. *P < 0.05, **P < 0.01 and ***P < 0.001. g, Fibronectin matrix changes from a relaxed framework to a rigid matrix. h, FSP1 expression in anterior (left) or posterior (right) regions of E12.5 ActinCre-ER;R26VT2/GK3 embryos after 24 h of 1 nM 4-hydroxytamoxifen exposure. Dotted lines delimitate single clones. i, Percentage of labelled cells in clones from two independent embryos. j, Local (subsampled) fractal analysis of a E16.5 En1Cre;R26VT2/GK3 dermis. Dashed lines delimitate dermal layers. k, Percentages ± s.e.m. of EPFs and ENFs in each layer; n = 3 optical fields. Two-way ANOVA, *P < 0.05, Tukey test. l, Mean ± s.e.m. number of cells per 1,000 µm2 of each layer; n = 3 optical fields. One-way ANOVA, *P < 0.05, Tukey test. m,n, FD values (m) and L values (n) derived from different dermal layers; n for FD and L, respectively = 58 and 58 (epidermis), 65 and 65 (DL-1), 198 and 199 (DL-2), 62 and 62 (DL-3), 274 and 281 (DL-4), 92 and 92 (DL-5), and 87 and 91 (hypodermis) subsampled values pooled from 3 confocal images. Box and whiskers plots with minimum, lower quartile, median, upper quartile and maximum. One-way ANOVA, *P < 0.05, Tukey test. Scale bars, 20 µm (c), 200 µm (h) and 50 µm (j). P values are listed in Supplementary Table 1.

Article Snippet: Antibodies used: Ki67 (Abcam), Cas3 (Cell Signaling), collagen I (Rockland), fibronectin (Abcam), FSP1 (Abcam), collagen III (Abcam), collagen VI (Abcam), CD31 (Novus Biologicals), α -SMA (Abcam), CD26 (Abcam), TNC (Abcam), Dlk1 (Abcam), Sca1 (Biolegend), FABP4 (Abcam) and AF647 secondary antibody (Molecular Probes).

Techniques: Derivative Assay, Expressing, Clone Assay

Journal: Nature cell biology

Article Title: Two succeeding fibroblastic lineages drive dermal development and the transition from regeneration to scarring.

doi: 10.1038/s41556-018-0073-8

Figure Lengend Snippet: Fig. 5 | Regeneration-to-scar transition is coupled to EPFs development. a, E12.5 En1Cre;R26mTmG embryos were collected, wounded on the back-skin and kept in culture for 48 h. b,c, Immunofluorescence for fibronectin (b) or collagen I (c). Dotted lines delimitate the lesion site. Arrows indicate EPFs. d,e, Representative immunofluorescence images of fibronectin staining at adjacent fetal skin (control) (d) and the wounded site (e) at 48 h post-wounding. f, FD values (left) and L values (right) derived from fibronectin staining at adjacent fetal skin (control) and the wounded site at 24 h and 48 h after wounding. n = 6 optical fields of control skins and n = 5 optical fields of wounds from two embryos. Mean ± s.e.m. Two-tailed unpaired Student’s t-test. g, Skin biopsies were taken from anterior, middle and posterior regions of E19.5 WT embryos and cultured for 5 days. h, Masson’s trichrome-stained sections of posterior or anterior biopsies. Arrows indicate the scar-like deposition of the ECM. i, FD values (left) and L values (right) derived from the cyan channel of Masson’s trichrome-stained biopsies. Anterior: n = 11, middle: n = 10 and posterior: n = 9. Mean ± s.e.m. One-way ANOVA, Tukey test, *P = 0.0436. Scale bars, 200 µm.

Article Snippet: Antibodies used: Ki67 (Abcam), Cas3 (Cell Signaling), collagen I (Rockland), fibronectin (Abcam), FSP1 (Abcam), collagen III (Abcam), collagen VI (Abcam), CD31 (Novus Biologicals), α -SMA (Abcam), CD26 (Abcam), TNC (Abcam), Dlk1 (Abcam), Sca1 (Biolegend), FABP4 (Abcam) and AF647 secondary antibody (Molecular Probes).

Techniques: Immunofluorescence, Staining, Control, Derivative Assay, Two Tailed Test, Cell Culture