Journal: Nature Communications

Article Title: Platelet PD-L1 reflects collective intratumoral PD-L1 expression and predicts immunotherapy response in non-small cell lung cancer

doi: 10.1038/s41467-021-27303-7

Figure Lengend Snippet: a , b Expression of GFP ( a ) and PD-L1 ( b ) on platelet surfaces after co-incubation with six FLAG-GFP- and PD-L1-GFP--transfected NSCLC cell lines (A549, NCI-H322, NCI-H522, NCI-H23, HOP-62, HOP-92) ( n = 3). c , d Ratio of GFP+ platelets/GFP+ tumor cells ( c ) and PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells ( d ) after co-incubation of platelets with transfected NSCLC cell lines ( n = 3). e Heat map of relative fibrinogen ( FBG ), tissue factor ( F3 ), fibronectin 1 ( FN1 ) and von Willebrand factor ( VWF ) mRNA levels in all tested NSCLC cell lines ( n = 3). f Relative mRNA level of FN1 in all tested NSCLC cell lines ( n = 3). g Correlation ratio of PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells and relative FN1 mRNA level ( n = 3). h Immunofluorescence images of platelet adhesion to NCI-H23 (upper left) and HOP-62 cells (upper right and lower) (fibronectin: green, platelets: red) ( n = 3). Upper scale bar 100 µm, lower 20 µm. i Left, Quantification of adhesive platelets after co-incubation with NCI-H23 and HOP-62. Quantified as CD61+ area in %/FoV ( n = 6 out of 3 independent experiments). Right, quantification of fibronectin covered area in %/FoV in NCI-H23 and HOP-62 cells ( n = 6 out of 3 independent experiments). j Correlation of platelet and fibronectin covered area in %/FoV. k Immunofluorescence images of PD-L1 and fibronectin expression in HOP-62 cells ( n = 2). Scale bar 20 µm. l Representative PLA with PD-L1 and fibronectin in HOP-62 cells. Scale bar left 20 µm, right 10 µm ( n = 1). m Representative PLA with PD-L1 and fibronectin in NCI-H23 and HOP-62 cell ( n = 3). Scale bar 10 µm. n PLA quantification of foci/cell in 119 NCI-H23 and 126 HOP-62 cells out of three biological replicates. o Western blot for PD-L1 and fibronectin in PD-L1-GFP-transfected HOP-62 cells after siRNA knockdown for fibronectin. Vinculin and α-Tubulin were used as loading controls ( n = 2). Presentation of full scan blots are provided in the Source data file. p Expression of PD-L1 on platelets after co-incubation with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). q Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP, PD-L1-GFP/siFN1, and PD-L1-GFP/siNC-transfected HOP-62 cells ( n = 3). r Representative images of platelet adhesion to fibronectin-coated surface in the presence or absence of different platelet-blocking agents ( n = 3). Scale bar 20 µm. s , t Quantitative analysis of the platelet adhesion assay as platelet covered area/FoV in % (s) and platelets/FoV (t) ( n = 9 (small symbols) were analyzed out of three independent experiments (large symbols)). Horizontal lines represent mean. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. u Quantification of PD-L1 on platelets after co-incubation with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). v Ratio PD-L1-GFP+ platelets/PD-L1-GFP+ tumor cells after co-incubation of platelets with PD-L1-GFP-transfected HOP-62 cells with or without pre-treatment with platelet-blocking agents ( n = 3). a – d , f , p , q , u , v Data are mean ± SEM. Statistical significance was calculated by one-way ANOVA and Tukey’s multiple comparisons test. g, j Correlation was determined by simple linear regression analysis. I, n Data are mean ± SEM. Statistical significance was calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.

Article Snippet: For siRNA knockdown of Fibronectin, Lipofectamine TM 3000 and 100 pmol of human (FN1) siRNA Oligo Duplex (Locus ID 2335) (Origene, SR320193) was used.

Techniques: Expressing, Incubation, Transfection, Immunofluorescence, Adhesive, Western Blot, Knockdown, Blocking Assay, Cell Adhesion Assay, Two Tailed Test

Journal: Epigenetics

Article Title: ANRIL upregulates TGFBR1 to promote idiopathic pulmonary fibrosis in TGF-β1-treated lung fibroblasts via sequestering let-7d-5p.

doi: 10.1080/15592294.2024.2435682

Figure Lengend Snippet: Figure 2. Knockdown of ANRIL inhibited TGF-b induced fibroblasts proliferation and activation. (a) The relative mRNA levels of Fn1, Col1a1, Col3a1, and Acta2 in ANRIL silenced MRC-5 cells after TGF-b induction; n = 3. (b) The relative protein levels of Fn1, Col1a1, Col3a1, and Acta2 in ANRIL silenced MRC-5 cells after TGF-β induction; n = 3. (c) EdU staining demonstrated the effect of ANRIL knockdown on lung fibroblast proliferation. Scale bar, 50 mm, n = 5. (d) Scratch assay for the evaluation of migration of lung fibroblasts after ANRIL knockdown. Scale bar, 200 mm, n = 5. (e) Immunofluorescence staining indicated that TGF-β1-induced a-sma positive cells were impeded by the reduced expression of ANRIL in MRC-5 fibroblasts. Scale bar, 50 mm, n = 5. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Antibodies against TGFBR1 (1:1000, Proteintech 30,117–1-AP), Acta2 (1:2000, CST, #14968), FN1 (1:2000, CST, #26836), Col1a1 (1:2000, CST, #72026), Col3a1 (1:2000, CST, #30565) were used to detect proteins.

Techniques: Knockdown, Activation Assay, Staining, Wound Healing Assay, Migration, Immunofluorescence, Expressing

Journal: Epigenetics

Article Title: ANRIL upregulates TGFBR1 to promote idiopathic pulmonary fibrosis in TGF-β1-treated lung fibroblasts via sequestering let-7d-5p.

doi: 10.1080/15592294.2024.2435682

Figure Lengend Snippet: Figure 6. ANRIL promotes lung fibroblasts activation via TGFBR1. (a,b). The relative mRNA levels of Fn1, Col1a1, Col3a1, and Acta2 in MRC- 5 cells with different treatments after TGF-b induction, n = 3. (c,d). EdU results show the proliferation and migration of MRC-5 fibroblasts after different treatments, scale bar, 50 mm, n = 5. (e,f). wound healing assays show the proliferation and migration of MRC-5 fibroblasts after different treatments, scale bar, 50 mm, n = 5. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Antibodies against TGFBR1 (1:1000, Proteintech 30,117–1-AP), Acta2 (1:2000, CST, #14968), FN1 (1:2000, CST, #26836), Col1a1 (1:2000, CST, #72026), Col3a1 (1:2000, CST, #30565) were used to detect proteins.

Techniques: Activation Assay, Migration

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: Knockout of GSDMD alleviated renal tubule morphological changes in OXO/adenine-induced mice and attenuated fibrosis (A and B) Histological changes in renal tubules after HE (A) and Masson (B) staining in different groups. Scale bar 50 μm and 100μm. (C and D) Western blot showing α-SMA levels in renal tissue from different groups (n = 6 animals per group). (C and E) Western blot showing E-cadherin levels in renal tissue from different groups (n = 5 animals per group). (F and G) RT‒PCR analysis of fibronectin (F) and collagen I (G) levels in renal tissue from different groups (n = 5 animals per group). (H) RT‒PCR analysis of IL-1β levels in renal tissue from different groups (n = 6 animals per group). Data for (D)-(H) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (D–H). WT, wild type. Norm-diet, normal diet. UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Knock-Out, Staining, Western Blot

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: The primer sequences used in RT‒PCR

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques:

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: GSDMD encouraged IL-1β expression and fibrosis in NRK-52E cells in vitro (A–D) HK-2 cells were stimulated with uric acid for 48 h. Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) levels in HK-2 cells in the different groups (n = 3 per group). (E–H) NRK-52E cells were stimulated with uric acid for 48 h. Western blot showing cleaved-caspase-1 (E and F), caspase-1 (E and G) and RIG-I (E and H) levels in NRK-52E cells in the different group (n = 6 per group). (I–P) NRK-52E cells were transfected with GSDMD-siRNA or NC-siRNA and treated with UA for 48 h. Western blot showing GSDMD (I and J) and GSDMD-N (I and K) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). (L) IL-1β level in culture medium (n = 6 per group). (M–P) Western blot showing α-SMA (M and N), E-cadherin (M and O) and fibronectin (M and P) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). Data for (B–D), (F–H), (J–L) and (N–P) are presented as mean ± SEM. Student’s t test for (B–D) and (F–H). One-way ANOVA with Student-Newman-Keuls test for (J–L) and (N–P). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Expressing, In Vitro, Western Blot, Transfection, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: RIG-I-siRNA decreased HUA-induced fibrosis and pyroptosis molecules expressions in NRK-52E cells in vitro NRK-52E cells were transfected with RIG-I-siRNA or NC-siRNA and treated with UA for 48 h. (A–D) Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (E–G) Western blot showing cleaved-caspase-1 (E and F) and GSDMD-N (E and G) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (H) IL-1β level in culture medium (n = 6 per group). (I–K) Western blot showing fibronectin (I and J) and α-SMA (I and K) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). Data for (B–D), (F–H) and (J–K) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (B–D), (F–H) and (J–K). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: In Vitro, Transfection, Western Blot, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet:

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Recombinant, H&E Stain, Staining, Activity Assay, Transfection, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Lysis, Bradford Protein Assay, Bicinchoninic Acid Protein Assay, Negative Control, Software, Fluorescence, Microscopy

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 2. Effects of A-485 on the profibrotic phenotype induced by TGF-β1 in HK-2 cells. (A) The representative western blot images and quantification of FN, Col-I, E- cad and α-SMA in cultured HK-2 cells (n = 3). (B) Micrographs of α-SMA and E-cad immunofluorescence staining in HK-2 cells (n = 6). (C-D) The quantification of α-SMA and E-cad immunofluorescence staining in HK-2 cells. Values are expressed as means ± SEM. *p < 0.05, * *p < 0.01. FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Cell Culture, Immunofluorescence, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 4. Effects of A-485 on UUO-induced profibrotic proteins in mouse kidney. (A-B) The representative western blot images and quantification of FN, Col-I, E-cad and α-SMA in mice (n = 6). (C) Representative micrographs of FN, Col-I and E-cad immunohistochemical staining in mice. (D-F) The quantification of FN, Col-I and E- cad expression detected by immunohistochemical staining (n = 5). Values are expressed as means ± SEM. * *p < 0.01. sham, sham operation; UUO: unilateral ureteral obstruction; FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Immunohistochemical staining, Staining, Expressing

Journal: Molecules

Article Title: Vincamine Ameliorates Epithelial-Mesenchymal Transition in Bleomycin-Induced Pulmonary Fibrosis in Rats; Targeting TGF-β/MAPK/Snai1 Pathway

doi: 10.3390/molecules28124665

Figure Lengend Snippet: Effect of vincamine on fibronectin ( A ), N-cadherin ( B ), and collagen ( C ) levels in lung tissues. Bars represent mean ± SD. Significant difference was analyzed by one-way ANOVA test followed by post hoc Dunnett test, where * p < 0.001, compared to sham group, and # p < 0.001, compared to BLM-induced group.

Article Snippet: In lung tissue homogenates, levels of fibronectin, N-cadherin, and collagen were also assessed, according to the manufacturer’s instructions, utilizing rat fibronectin ELISA kit (#MBS761397, MyBioSource, CA, USA), N-cadherin ELISA kit (#KOA0665, Rockland immunochemical Inc., Royersford, PA, USA), and collagen type I ELISA kit (#MBS262647, MyBioSource, CA, USA), respectively.

Techniques:

Journal: Pharmacological Reports

Article Title: WIN55,212-2 attenuates intestinal fibrosis in a DSS-induced mouse model

doi: 10.1007/s43440-026-00822-0

Figure Lengend Snippet: The effect of CB1 agonist (WIN 55,212-5; 5 mg/kg, 100 µL/animal, ip, once daily on days 10–78 from induction of colitis) in the chronic mouse model of colitis induced by dextran sulfate sodium (DSS) on relative mRNA expression of ( A ) alpha smooth muscle actin 2 (Acta2), ( B ) collagen type 1 alpha (Col1a1), ( C ) fibronectin 1 (Fn1), ( D ) SMAD family member 3 (Smad3). Kruskal-Wallis test and post hoc Dunn’s test were applied in all measurements. Real-time PCR was used for gene expression analysis. Data represent median and min-max and range of 5–7 mice per group

Article Snippet: Ultimate stage was the quantification of mRNA expression, which was performed using real-time fluorescence detection PCR technique with the use of fluorescently labeled TaqMan probes (Thermofisher, Waltham, USA): mouse probes for Acta2 (Mm00725412_s1), Col1a1 (Mm00801666_g1), Smad3 (Mm01170760_m1) and Fn1 (Mm01256744_m1) and human probes for Acta2 (Hs00426835_g1), COL1A1 (Hs00164004_m1), Smad3 (Hs00969210_m1) and Fn1 (Hs01549976_m1).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Pharmacological Reports

Article Title: WIN55,212-2 attenuates intestinal fibrosis in a DSS-induced mouse model

doi: 10.1007/s43440-026-00822-0

Figure Lengend Snippet: Relative mRNA expression of ( A ) alpha smooth muscle actin 2 (ACTA2), ( B ) collagen type 1 alpha 1 (COL1A1), ( C ) fibronectin 1 (FN1), ( D ) SMAD family member 3 (SMAD3) in human samples: controls (HC) and ulcerative colitis patients (UC). The U-Mann test was applied in all measurements. Real-time PCR was used for gene expression analysis. Data represent median and min-max range. **** - p < 0.0001

Article Snippet: Ultimate stage was the quantification of mRNA expression, which was performed using real-time fluorescence detection PCR technique with the use of fluorescently labeled TaqMan probes (Thermofisher, Waltham, USA): mouse probes for Acta2 (Mm00725412_s1), Col1a1 (Mm00801666_g1), Smad3 (Mm01170760_m1) and Fn1 (Mm01256744_m1) and human probes for Acta2 (Hs00426835_g1), COL1A1 (Hs00164004_m1), Smad3 (Hs00969210_m1) and Fn1 (Hs01549976_m1).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/elife.58688

Figure Lengend Snippet: Figure 5. Impact of LOX inhibition on T cell migration in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Migration of activated PBT plated onto fresh tumor slices was analyzed in EGI-1 and MMTV-PyMT tumor model, whilst resident tumor-infiltrating T lymphocytes were analyzed in mPDAC and KPC tumor model. Illustrative images of T cell migration tracks in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Tumor stroma (fibronectin) in red, tumor cells (EpCAM in EGI-1, MMTV-PyMT, and KPC tumor models, CD44 in mPDAC tumor models), in blue and T cells (CD8 in mPDAC and KPC, Calcein in MMTV-PyMT, and EGI-1 tumor models) in green. Tracks are color-coded to illustrate track displacement. Scale bar = 100 mm. T cell migration speed, T cell displacement, and trajectory straightness in all tumor models. ***p-value>0.001, p-value>0.05, Student’s t-test. Results are shown as mean ± SD. The online version of this article includes the following source data and figure supplement(s) for figure 5:

Article Snippet: The slices were then washed to remove all cells that had not infiltrated the slice and stained for 15 min at 37 ̊C with the following antibodies: BV421–anti- human EpCAM (9C4 clone; BioLegend) and anti-human/mouse fibronectin at 10 mg/mL (HFN7.1 clone; Acris antibodies).

Techniques: Inhibition, Migration

Journal: Renal Failure

Article Title: apoE Expression in Glomerulus and Correlation with Glomerulosclerosis Induced by Adriamycin in Rats

doi: 10.3109/0886022x.2011.559680

Figure Lengend Snippet: Figure 1. Glomerular morphology was normal in sham group (A1: 9 weeks; A2: 13 weeks; HE). Proliferation occurring in the majority of glomerular mesangial cells and extracellular matrix in GS group, and degeneration of glomerular epithelial cells and infiltration of wide- spread mononuclear cells were shown (A3: 9 weeks; A4: 13 weeks; HE), especially in 13 weeks of GS group (A4). Representative samples of immunohistochemical staining for glomerular Col-IV (SHO: B1: 9 weeks, B2: 13 weeks; GS: B3: 9 weeks and B4: 13 weeks), FN (SHO: C1: 9 weeks, C2: 13 weeks; GS: C3: 9 weeks and C4: 13 weeks), a-SMA (SHO: D1: 9 weeks, D2: 13 weeks; GS: D3: 9 weeks and D4: 13 weeks), TGF-b1 (SHO: E1: 9 weeks, E2: 13 weeks; GS: E3: 9 weeks and E4: 13 weeks), and apoE (SHO: F1: 9 weeks, F2: 13 weeks; GS: F3: 9 weeks and F4: 13 weeks) were observed in all groups. Sham group (B1, B2, C1, C2, D1, D2, E1, E2, F1, and F2): positive staining (in brown) was faint in glomerulus, glomerular endothelial cells, glomerular basement membrane, mesangial cells, and visceral epithelial cells. GS group (B3, B4, C3, C4, D3, D4, E3, E4, F3, and F4): positive staining was strong in most glomerulus, glomerular endothelial cells, glomerular basement membrane, mesangial cells, and visceral epithelial cells, especially in those of 13 weeks (B4, C4, D4, E4, and F4). Magnification 400×. Notes: SHO, sham operation group; GS, glomerulosclerosis model group; HE, hematoxylin and eosin; FN, fibronectin; a-SMA, a-smooth muscle actin; TGF-b1, transforming growth factor-b1; apoE, apolipoprotein E.

Article Snippet: Finally, the sections were incubated with primary monoclonal antibodies against apoE (1/150) (Bo Ao-Sen, Co., Beijing, China), Collagen IV (Col-IV; 1/100) (Shanghai ChangDao Co., Shanghai, China), fibronectin (FN; 1/ 100) (Beijing Zhongshan Co. Ltd., Beijing, China), a-smooth muscle actin (a-SMA) (ready-to-use kit) (Shanghai Changdao, Co.), and transforming growth factor-b1 (TGF-b1; 1/100) (Wuhan Boshide, Co., Wuhan, China) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining, Membrane

Journal: Oncotarget

Article Title: Snail and serpinA1 promote tumor progression and predict prognosis in colorectal cancer.

doi: 10.18632/oncotarget.3964

Figure Lengend Snippet: Figure 5: SerpinA1 was regulated by Snail. A. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail and serpinA1 protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (serpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and western blot analysis was performed for detection of Snail and SerpinA1 expression. C. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail) or control vector pcDNA (vector), and ChIP assays were performed. The presence of the serpinA1 promoter (-516/-4) was verified in immunoprecipitates with either mouse IgG or anti-Snail antibodies, and assay inputs were analyzed using real-time PCR. The samples were loaded on agarose gels. D. Data show promoter enrichment in the anti- Snail immunoprecipitate relative to IgG.

Article Snippet: For overexpression of genes, cells were grown to 60–70% confluence and transfected with the pcDNA Snail-Myc, pcDNA SerpinA1-Myc vectors or pcDNA fibronectin-Myc vectors (Origene; MD, USA), or with the pcDNA-3.1 vector as a control, with Lipofectamine 2000 (Invitrogen; Life Technologies; NY, USA) according to the manufacturer’s protocol.

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Snail and serpinA1 promote tumor progression and predict prognosis in colorectal cancer.

doi: 10.18632/oncotarget.3964

Figure Lengend Snippet: Figure 6: Snail and serpinA1 promoted tumor progression through fibronectin. A. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (SerpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. C. DLD-1 and SW480 cells were transfected with pcDNA-fibronectin (Fibronectin), control vector pcDNA (vector), fibronectin siRNA (siFibronectin), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. D., E. Invasion and migration assays were performed using transfected cells. Representative data are shown for cells that invaded (top) and migrated (bottom) in the presence of 1% FBS. *P < 0.05.

Article Snippet: For overexpression of genes, cells were grown to 60–70% confluence and transfected with the pcDNA Snail-Myc, pcDNA SerpinA1-Myc vectors or pcDNA fibronectin-Myc vectors (Origene; MD, USA), or with the pcDNA-3.1 vector as a control, with Lipofectamine 2000 (Invitrogen; Life Technologies; NY, USA) according to the manufacturer’s protocol.

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Migration

Journal: Oncotarget

Article Title: Snail and serpinA1 promote tumor progression and predict prognosis in colorectal cancer.

doi: 10.18632/oncotarget.3964

Figure Lengend Snippet: Figure 7: The roles of Snail and serpinA1 in other human cancer cell lines. A., B. MCF7, MDA-MB-231, A2780, and SKVO3 cells were transfected with pcDNA-Snail, pcDNA-serpinA1, Snail siRNA, or serpinA1 siRNA, and migration assays were performed using these cells. Graphs show the number of cells that migrated in the presence or absence of 1% FBS. *P < 0.05. C. MCF7, MDA- MB-231, A2780, and SKVO3 cells cells were transfected with pcDNA-Snail (Snail) or control vector pcDNA (vector), and ChIP assays were performed. The presence of the serpinA1 promoter (-516/-4) was verified in immunoprecipitates with either mouse IgG or anti-Snail antibodies, and assay inputs were analyzed using real-time PCR. The samples were loaded on agarose gels. D. Data shows promoter enrichment in the anti-Snail immunoprecipitate relative to IgG.

Article Snippet: For overexpression of genes, cells were grown to 60–70% confluence and transfected with the pcDNA Snail-Myc, pcDNA SerpinA1-Myc vectors or pcDNA fibronectin-Myc vectors (Origene; MD, USA), or with the pcDNA-3.1 vector as a control, with Lipofectamine 2000 (Invitrogen; Life Technologies; NY, USA) according to the manufacturer’s protocol.

Techniques: Transfection, Migration, Control, Plasmid Preparation, Real-time Polymerase Chain Reaction