Journal: Antioxidants

Article Title: Hippuric Acid Promotes Renal Fibrosis by Disrupting Redox Homeostasis via Facilitation of NRF2–KEAP1–CUL3 Interactions in Chronic Kidney Disease

doi: 10.3390/antiox9090783

Figure Lengend Snippet: Effects of hippuric acid (HA) on fibrotic gene expression in HK-2 cells. ( A ) Relative mRNA and ( B ) protein expression levels of collagen-I (COL1A1), vimentin (VIM), E-cadherin (CDH1), alpha-smooth muscle actin (ACTA2), matrix metalloproteinase-9 (MMP9), and tissue inhibitor of metalloproteinases-1 (TIMP1) after treatment with 0–1000 μM HA. Quantification was performed by densitometry. ( C ) Immunofluorescence staining for CDH1 (red) and ACTA2 (green) after treatment with 250 and 1000 μM HA. Nuclei (blue) were stained with DAPI. Bar = 100 μm. Data are presented as the mean ± standard deviation. * p < 0.05 vs. control (Ctrl) cells. Experiments were repeated three to four times.

Article Snippet: Antibodies containing collagen-I (COL1A1), E-cadherin (CDH1), alpha-smooth muscle actin (ACTA2), vimentin (VIM), heme oxygenase-1 (HO1), KEAP1, SNAI1, matrix metalloproteinase-9 (MMP9), NAD(P)H quinone dehydrogenase (NQO1), β-ACTIN, histone H3, and secondary antibodies were purchased from Proteintech (Chicago, IL, USA).

Techniques: Gene Expression, Expressing, Immunofluorescence, Staining, Standard Deviation, Control

Journal: Antioxidants

Article Title: Hippuric Acid Promotes Renal Fibrosis by Disrupting Redox Homeostasis via Facilitation of NRF2–KEAP1–CUL3 Interactions in Chronic Kidney Disease

doi: 10.3390/antiox9090783

Figure Lengend Snippet: Role of oxidative stress in hippuric acid (HA)-induced fibrotic gene expression in HK-2 cells. ( A ) Levels of reactive oxygen species (ROS) and hydrogen peroxide (H 2 O 2 ) after treatment with 0–1000 μM HA. ( B ) Relative mRNA and protein expression levels of NADPH oxidase 4 (NOX4) after treatment with 0–1000 μM HA. ( C ) Levels of protein expression of collagen-I (COL1A1), vimentin (VIM), E-cadherin (CDH1), and alpha-smooth muscle actin (ACTA2) in cells pretreated with 1000 µM N -acetylcysteine (NAC), 50 µM diphenylene iodonium (DPI), or the vehicle, followed by treatment with 1000 µM HA. ( D ) Immunofluorescence staining for CDH1 (red) and ACTA2 (green) after pretreatment with 1000 µM NAC, 50 µM DPI, or the vehicle, followed by treatment with 1000 µM HA. Nuclei (blue) were stained with DAPI. Bar = 100 μm. ( E ) Expression levels of key proteins in the TGFβ/SMAD signaling pathway in cells pretreated with 1000 µM NAC, 50 µM DPI, or the vehicle, followed by treatment with 1000 µM HA. Quantification was performed by densitometry. Data are presented as the mean ± standard deviation. * p < 0.05 vs. control (Ctrl) cells; # p < 0.05 vs. cells treated with HA only. Experiments were repeated three to five times.

Article Snippet: Antibodies containing collagen-I (COL1A1), E-cadherin (CDH1), alpha-smooth muscle actin (ACTA2), vimentin (VIM), heme oxygenase-1 (HO1), KEAP1, SNAI1, matrix metalloproteinase-9 (MMP9), NAD(P)H quinone dehydrogenase (NQO1), β-ACTIN, histone H3, and secondary antibodies were purchased from Proteintech (Chicago, IL, USA).

Techniques: Gene Expression, Expressing, Immunofluorescence, Staining, Standard Deviation, Control

Journal: Antioxidants

Article Title: Hippuric Acid Promotes Renal Fibrosis by Disrupting Redox Homeostasis via Facilitation of NRF2–KEAP1–CUL3 Interactions in Chronic Kidney Disease

doi: 10.3390/antiox9090783

Figure Lengend Snippet: Role of nuclear factor erythroid 2-related factor 2 (NRF2) in hippuric acid (HA)-induced fibrotic gene expression in HK-2 cells. ( A ) Levels of protein expression of NRF2, heme oxygenase-1 (HO1), and NAD(P)H quinone dehydrogenase (NQO1) in cells pretreated with 0–20 µM sulforaphane (SFN), followed by treatment with 1000 µM HA. ( B ) Levels of protein expression of collagen-I (COL1A1), vimentin (VIM), E-cadherin (CDH1), and alpha-smooth muscle actin (ACTA2) in cells pretreated with 0–20 µM SFN, followed by treatment with 1000 µM HA. ( C ) Levels of protein expression of NRF2, HO1, and NQO1 after cell transfection with the control or NRF2 -specific small interfering RNA (siRNA), followed by treatment with 1000 µM HA. ( D ) Levels of protein expression of COL1A1, VIM, CDH1, and ACTA2 after cell transfection with the control or NRF2 -specific siRNA, followed by treatment with 1000 µM HA. Quantification was performed by densitometry. Data are presented as the mean ± standard deviation. * p < 0.05 vs. control (Ctrl) cells; # p < 0.05 vs. cells treated with HA only or HA + control siRNA. Experiments were repeated three times.

Article Snippet: Antibodies containing collagen-I (COL1A1), E-cadherin (CDH1), alpha-smooth muscle actin (ACTA2), vimentin (VIM), heme oxygenase-1 (HO1), KEAP1, SNAI1, matrix metalloproteinase-9 (MMP9), NAD(P)H quinone dehydrogenase (NQO1), β-ACTIN, histone H3, and secondary antibodies were purchased from Proteintech (Chicago, IL, USA).

Techniques: Gene Expression, Expressing, Transfection, Control, Small Interfering RNA, Standard Deviation

Journal: International Journal of Oncology

Article Title: E2F1/SP3/STAT6 axis is required for IL-4-induced epithelial-mesenchymal transition of colorectal cancer cells

doi: 10.3892/ijo.2018.4429

Figure Lengend Snippet: IL-4-induced epithelial-mesenchymal transition program is more evident in HCT116 cells compared with in RKO cells. (A) Left: Representative images of IL-4 immunostaining in human colorectal cancer tissue and para-tumor tissue. Scale bar, 200 μ m. Right: Immunostaining scores of IL-4 in invasive or non-invasive CRC tissues. (B) Morphology of HCT116 and RKO cells treated with or without IL-4 (20 ng/ml) for 72 h visualized using phase-contrast microscopy. Scale bar, 500 μ m. (C) Left: Representative images from wound healing assays with HCT116 and RKO cells treated with 20 ng/ml IL-4. Scale bar, 100 μ m. Right: Percentage wound closure 48 h after addition of IL-4. (D) Left: Representative images of HCT116 and RKO cells penetrating the Matrigel in invasion assays following treatment with 20 ng/ml IL-4. Scale bar, 100 μ m. Right: numbers of invasive cells treated with IL-4 for 48 h. (E) Relative mRNA levels of vimentin, fibronectin, E-cadherin and ZO-1 in HCT116 (left) and RKO (right) cells treated with 20 ng/ml IL-4 for 48 h. (F) Western blots of vimentin, fibronectin, E-cadherin and ZO-1 with specific antibodies in HCT116 (left) and RKO (right) cells treated with 20 ng/ml IL-4 for 48 h. * P<0.05; ** P<0.01. IL, interleukin; CRC, colorectal cancer; E-cadherin, epithelial cadherin; ZO-1, zonula occludens-1.

Article Snippet: Antibodies against E-cadherin (cat. no. sc-7870; 1:1,000), vimentin (cat. no. sc-6260; 1:1,000), E2F1 (cat. no. sc-193; 1:1,000) and GAPDH (cat. no. sc-365062; 1:5,000) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Immunostaining, Microscopy, Western Blot

Journal: International Journal of Oncology

Article Title: E2F1/SP3/STAT6 axis is required for IL-4-induced epithelial-mesenchymal transition of colorectal cancer cells

doi: 10.3892/ijo.2018.4429

Figure Lengend Snippet: Highly expressed E2F1 promotes IL-4-induced aggressiveness by upregulating STAT6. (A) Western blots of the indicated proteins with specific antibodies in HCT116 cells co-transfected with E2F1 siRNA/STAT6 overexpression plasmid and exposed to 20 ng/ml IL-4 for 48 h. (B) Left: Representative images from wound healing and invasion assays with HCT116 cells co-transfected with E2F1 siRNA/STAT6 overexpression plasmid and exposed to 20 ng/ml IL-4 for 48 h. Scale bar, 100 μ m. Right: Percentage wound closure and numbers of invasive cells 48 h after IL-4 addition. (C) Western blots of the indicated proteins with specific antibodies in the RKO cells co-transfected with STAT6 siRNA/E2F1 overexpression plasmid and exposed to 20 ng/ml IL-4 for 48 h. (D) Left: representative images from wound healing and invasion assays with the RKO cells co-transfected with STAT6 siRNA/E2F1 overexpression plasmid and exposed to 20 ng/ml IL-4 for 48 h. Scale bar, 100 μ m. Right: Percentage wound closure and numbers of invasive cells 48 h after IL-4 addition. * P<0.05. IL, interleukin; STAT6, signal transducer and activator of transcription 6; si, small interfering RNA; Ctrl, control; EV, empty vector; E-cadherin, epithelial cadherin; ZO-1, zonula occludens-1.

Article Snippet: Antibodies against E-cadherin (cat. no. sc-7870; 1:1,000), vimentin (cat. no. sc-6260; 1:1,000), E2F1 (cat. no. sc-193; 1:1,000) and GAPDH (cat. no. sc-365062; 1:5,000) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Western Blot, Transfection, Over Expression, Plasmid Preparation, Small Interfering RNA, Control

Journal: International Journal of Oncology

Article Title: E2F1/SP3/STAT6 axis is required for IL-4-induced epithelial-mesenchymal transition of colorectal cancer cells

doi: 10.3892/ijo.2018.4429

Figure Lengend Snippet: As a target gene of STAT6, Zeb1 is important in EMT triggered by IL-4. (A) mRNA levels of EMT transcription factors in HCT116 cells treated with 20 ng/ml IL-4 for 24 h. (B) Western blots of the indicated proteins with specific antibodies in HCT116 cells treated with 20 ng/ml IL-4 for 24 h. (C) mRNA levels of Zeb1/2 in HCT116 cells with STAT6-knockdown. (D) Western blots of the indicated proteins with specific antibodies in HCT116 cells transfected with STAT6 siRNA and exposed to 20 ng/ml IL-4 for 24 h. (E) Western blots of the indicated proteins with specific antibodies in HCT116 cells co-transfected with STAT6 siRNA/Zeb1 overexpression plasmid and exposed to 20 ng/ml IL-4 for 48 h. * P<0.05; ** P<0.01. STAT6, signal transducer and activator of transcription 6; EMT, epithelial-mesenchymal transition; IL, interleukin; Zeb, zinc finger E-box-binding homeobox; si, small interfering; Ctrl, control; E-cadherin, epithelial cadherin; ZO-1, zonula occludens-1; EV, empty vector.

Article Snippet: Antibodies against E-cadherin (cat. no. sc-7870; 1:1,000), vimentin (cat. no. sc-6260; 1:1,000), E2F1 (cat. no. sc-193; 1:1,000) and GAPDH (cat. no. sc-365062; 1:5,000) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Western Blot, Knockdown, Transfection, Over Expression, Plasmid Preparation, Binding Assay, Control

Journal: Oncology Letters

Article Title: Decreased expression of CD63 tetraspanin protein predicts elevated malignant potential in human esophageal cancer

doi: 10.3892/ol.2017.6023

Figure Lengend Snippet: Western blot analysis of the expression of E-cadherin, vimentin, MMP-2 and MMP-9 following treatment with small interfering RNA targeting CD63. The epithelial marker E-Cadherin was downregulated and the mesenchymal maker vimentin was upregulated by CD63 knockdown. MMP-2 and MMP-9 expression levels were increased by CD63 knockdown. MMP, matrix metalloproteinase; NC, negative control; siCD63, small interfering RNA targeting CD63.

Article Snippet: Following blocking with 5% nonfat milk in TBST (20 mM Tris, pH 7.5; 500 mM NaCl; 0.05% Tween-20) for 1 h at room temperature, the membranes were then incubated overnight at 4°C with the primary antibody to CD63 (cat. no., ab134045; dilution, 1:1,000, Abcam), E-cadherin (cat. no., sc-59778; dilution, 1:200, Santa Cruz Biotechnology), vimentin (cat. no. sc-6260; dilution, 1:200, Santa Cruz Biotechnology), MMP-2 (cat. no. sc-13594; dilution, 1:200, Santa Cruz Biotechnology) and MMP-9 (cat. no. sc-21733; dilution, 1:200, Santa Cruz Biotechnology). β-Actin (cat. no. sc-8432; dilution, 1:200, Santa Cruz Biotechnology) was used as a loading control.

Techniques: Western Blot, Expressing, Small Interfering RNA, Marker, Knockdown, Negative Control

Journal: Blood Advances

Article Title: Plasma kallikrein supports FXII-independent thrombin generation in mouse whole blood

doi: 10.1182/bloodadvances.2024012613

Figure Lengend Snippet: Effect of PKa inhibition on contact pathway–initiated TG in FXII-sufficient and -deficient WB. Representative curves of silica-initiated TG in F12 +/+ (A) and F12 – / – (B) mouse WB in the presence of increasing concentrations of the PKa inhibitor (PKai) berotralstat or vehicle control. Quantification of TG lag time (C) and peak TG (D) in F12 +/+ or F12 – / – WB in the presence of 8 μM berotralstat or vehicle control (n = 3 per group). (E) Representative curves of silica-initiated TG in F12 – / – mouse WB in the presence of increasing concentrations of the inhibitory anti-PKa antibody lanadelumab or IgG control. Quantification of TG lag time (F) and peak TG (G) in F12 – / – WB in the presence of 400 μg/mL lanadelumab or IgG control (n = 3 per group). Data presented as individual values with mean ± standard deviation (SD). Data analyzed with paired Student t tests comparing samples with and without inhibitor; ∗ P < .05; ∗∗ P < .01. Bero, berotralstat; Lana, lanadelumab; ns, not significant; Veh, vehicle.

Article Snippet: In some experiments, samples were supplemented with the small molecule PKa inhibitor berotralstat (MedChem Express, Monmouth Junction, NJ), an anti-FXIIa antibody (garadacimab/CSL-312 bio-similar; ATUM Bio, Newark, CA), an anti-PKa antibody (lanadelumab bio-similar; ProteoGenix USA, Miami, FL), an anti-TF antibody (1H1; Genentech, San Francisco, CA), or appropriate vehicle and immnuoglubulin G (IgG) controls.

Techniques: Inhibition, Control, Standard Deviation

Journal: Blood Advances

Article Title: Plasma kallikrein supports FXII-independent thrombin generation in mouse whole blood

doi: 10.1182/bloodadvances.2024012613

Figure Lengend Snippet: Effect of PKa inhibition on contact pathway–initiated TG in FXII-deficient PRP and RBC-reconstituted PRP. Representative curves of silica-initiated TG in F12 – / – PRP (A) and F12 – / – PRP reconstituted with RBCs (PRP+RBCs) (B) in the presence of the PKa inhibitor berotralstat or vehicle control. Quantification of TG lag time (C) and peak TG (D) in F12 – / – PRP and PRP with RBCs (n = 4 per group). Data presented as mean ± SD with paired t tests comparing samples with and without inhibitor; ∗∗ P < .01; ∗∗∗ P < .001. Bero, berotralstat; ns, not significant; Veh, vehicle.

Article Snippet: In some experiments, samples were supplemented with the small molecule PKa inhibitor berotralstat (MedChem Express, Monmouth Junction, NJ), an anti-FXIIa antibody (garadacimab/CSL-312 bio-similar; ATUM Bio, Newark, CA), an anti-PKa antibody (lanadelumab bio-similar; ProteoGenix USA, Miami, FL), an anti-TF antibody (1H1; Genentech, San Francisco, CA), or appropriate vehicle and immnuoglubulin G (IgG) controls.

Techniques: Inhibition, Control

Journal: Blood Advances

Article Title: Plasma kallikrein supports FXII-independent thrombin generation in mouse whole blood

doi: 10.1182/bloodadvances.2024012613

Figure Lengend Snippet: Effect of endogenous TF on contact pathway–initiated TG in WB. Representative curves of silica-initiated TG in TF low or TF wt mouse WB (A) and TF low or TF wt mouse WB (B) in the presence of 8 μM berotralstat and 50 μg/mL garadacimab with quantification of TG lag time (C) and peak TG (n = 4-5 per group) (D). Data presented as individual values with mean ± SD. Data analyzed with paired Student t tests comparing TF low and TF wt ; ∗∗ P < .01. Bero, berotralstat; ns, not significant; Veh, vehicle.

Article Snippet: In some experiments, samples were supplemented with the small molecule PKa inhibitor berotralstat (MedChem Express, Monmouth Junction, NJ), an anti-FXIIa antibody (garadacimab/CSL-312 bio-similar; ATUM Bio, Newark, CA), an anti-PKa antibody (lanadelumab bio-similar; ProteoGenix USA, Miami, FL), an anti-TF antibody (1H1; Genentech, San Francisco, CA), or appropriate vehicle and immnuoglubulin G (IgG) controls.

Techniques:

Journal: Cellular Oncology

Article Title: Upregulation of limb-bud and heart (LBH) drives liver cancer progression by interacting with the oncoprotein Mortalin

doi: 10.1007/s13402-026-01228-z

Figure Lengend Snippet: LBH is transcriptionally upregulated by TGF-β in liver cancer cells. (A) Heatmap displaying the common 474 DEGs whose expressions were altered upon treatment with 100 pM of TGF-β1 for 4 h in three different HCC cell lines, i.e. MHCC97H, MHCC97L and HCCLM3. (B) Heatmap classification displaying some typical upregulated genes in response to TGF-β1 administration in HCC cell lines. (C-H) After being treated with 100 pM of TGF-β1 (C-E) or 10 µM of SB431542 (F-H) for the indicated periods, HCCLM3, HLE and SMMC-7721 cells were harvested for RNA purification and gene expression analyses through qPCR. (I-J) HCCLM3, HLE cells transfected with a control siRNA (NS) or those targeting Smad2/3 were treated with 100 pM of TGF-β1 for 6 h. Then the expression levels of Smad2, Smad3 and LBH were analyzed by qPCR. (K) Gene track view of Smad2/3, H3K4me3, and H3K27ac at the LBH promoter from the Cistrome Data database (top), and schematic diagram of ChIP primers designed within and near the LBH promoter region (bottom). HCCLM3 cells were treated with 2.5 ng/ml TGF-β for 24 h. ChIP assays were then performed to examine the binding of Smad2/3 to the LBH promoter region, and the enriched DNA fragments were quantitatively analyzed by qPCR. qPCR experiments described above were analyzed by one-way ANOVA followed by Tukey’s or Dunnett’s post hoc test. Statistical significance: * p < 0.05, ** p < 0.01 and *** p < 0.001; ns, no significance. Data were shown as mean ± SD

Article Snippet: Recombinant human TGF-β1 protein was obtained from R&D Systems Inc., and the small molecule inhibitor SB431542 was purchased from MedChemExpress (MCE, USA).

Techniques: Purification, Gene Expression, Transfection, Control, Expressing, Binding Assay

Journal: Experimental and Therapeutic Medicine

Article Title: ASPM combined with KIF11 promotes the malignant progression of hepatocellular carcinoma via the Wnt/β-catenin signaling pathway

doi: 10.3892/etm.2021.10588

Figure Lengend Snippet: Sequences of the primers used for reverse transcription-quantitative PCR.

Article Snippet: N-cadherin , 1:5,000 , ab76011 , Rabbit monoclonal , Abcam.

Techniques: Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: ASPM combined with KIF11 promotes the malignant progression of hepatocellular carcinoma via the Wnt/β-catenin signaling pathway

doi: 10.3892/etm.2021.10588

Figure Lengend Snippet: ASPM-knockdown inhibits the migration, invasion and epithelial-to-mesenchymal transition of hepatocellular carcinoma cells. (A) Migration was detected using a wound healing assay; magnification, x100. (B) Invasion was detected using a Transwell assay; magnification, x100. (C) Protein expression levels of MMP2 and MMP9 were detected by western blotting. Expression levels of N-cadherin, vimentin and E-cadherin were detected by (D) reverse transcription-quantitative PCR and (E) western blotting. *** P<0.001; n≥3. ASPM, assembly factor for spindle microtubules; si, small interfering RNA; NC, negative control.

Article Snippet: N-cadherin , 1:5,000 , ab76011 , Rabbit monoclonal , Abcam.

Techniques: Knockdown, Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: ASPM combined with KIF11 promotes the malignant progression of hepatocellular carcinoma via the Wnt/β-catenin signaling pathway

doi: 10.3892/etm.2021.10588

Figure Lengend Snippet: ASPM-knockdown inhibits the migration, invasion and epithelial-to-mesenchymal transition of hepatocellular cancer cells via KIF11. (A) Migration was detected using a wound healing assay; magnification, x100. (B) Invasion was detected using a Transwell assay; magnification, x100. (C) Protein expression levels of MMP2 and MMP9 were detected by western blotting. Expression levels of N-cadherin, vimentin and E-cadherin were detected by (D) reverse transcription-quantitative PCR and (E) western blotting. ** P<0.01; *** P<0.001; n≥3. ASPM, assembly factor for spindle microtubules; KIF11, kinesin family member 11; si, small interfering RNA; Ov, overexpression; NC, negative control.

Article Snippet: N-cadherin , 1:5,000 , ab76011 , Rabbit monoclonal , Abcam.

Techniques: Knockdown, Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA, Over Expression, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: miR-488-3p overexpression inhibits LSCC cell proliferation, migration, invasion and epithelial-mesenchymal transition. (A) Quantitative PCR analysis of miR-488-3p expression in LSCC cells transfected with miR-488-3p or NC mimics. (B) Cell Counting Kit-8 and (C) colony formation assays detected the proliferative ability of LSCC cells transfected with miR-488-3p or NC mimics. Transwell assays determined the (D) migration and (E) invasion of LSCC cells transfected with miR-488-3p or NC mimics. (F) Western blotting of the protein levels of E-cadherin, N-cadherin and vimentin. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. Scale bars, 100 µ m. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; LSCC, laryngeal squamous cell carcinoma; NC, negative control.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: Over Expression, Migration, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: ACVR1C knockdown inhibits LSCC cell proliferation, migration, invasion and epithelial-mesenchymal transition. (A) Analysis of ACVR1C expression in RNA-seq data ( GSE127165 ). (B) Quantitative PCR and (C) western blot analysis of the mRNA and protein expression levels of ACVR1C in AMC-HN-8 and FD-LSC-1 cells transfected with si-ACVR1C-1 and 2 or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. (D) Cell Counting Kit-8 and (E) colony formation assays investigated the proliferative ability of LSCC cells transfected with si-ACVR1C or si-NC. In the Transwell assay, ACVR1C knockdown significantly suppressed the (F) migration and (G) invasion of LSCC cells. (H) Western blot analysis of the protein levels of E-cadherin, N-cadherin and vimentin in LSCC cells transfected with si-ACVR1C or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. Scale bar, 100 µ m. * P<0.05, ** P<0.01 and *** P<0.001. ACVR1C, activin A receptor type 1C; LSCC, laryngeal squamous cell carcinoma; NC, negative control; si, small interfering RNA.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: Knockdown, Migration, Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Cell Counting, Transwell Assay, Negative Control, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: Effect of miR-488-3p on tumor growth in vivo . (A) Image of xenograft tumors after intratumoral injection of miR-488-3p or NC agomir. The xenograft tumor volume was measured to plot the growth curve. (B) Final weight of xenograft tumors. (C) Quantitative PCR analysis of miR-488-3p expression in xenograft tumors. (D) Representative immunohistochemical staining of vimentin, N-cadherin, E-cadherin and Ki-67 expression in xenograft tumors. (E) Schematic representation of the MEIS1/miR-488-3p/ACVR1C axis in regulating laryngeal squamous cell carcinoma progression. The red and green arrows indicate upregulation and downregulation in LSCC, respectively. Scale bar, 50 mm. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; NC, negative control; MEIS1, myeloid ecotropic viral integration site 1; ACVR1C, activin A receptor type 1C.

Article Snippet: The primary antibodies used were as follows: ACVR1C (1:1,000; cat. no. NBP1-50659; Novus Biologicals, LLC), N-cadherin (1:1,000; cat. no. 13116S; Cell Signaling Technology, Inc.), E-cadherin (1:1,000; cat. no. 3195S; Cell Signaling Technology, Inc.), vimentin (1:1,000; cat. no. 5741S; Cell Signaling Technology, Inc.), Flag (1:1,000; cat. no. F1804; Sigma-Aldrich; Merck KGaA) and GAPDH (1:1,000; cat. no. HC301-02; TransGen Biotech Co., Ltd.).

Techniques: In Vivo, Injection, Real-time Polymerase Chain Reaction, Expressing, Immunohistochemical staining, Staining, Negative Control

Journal: iScience

Article Title: Targeting ITGB4/SOX2-driven lung cancer stem cells using proteasome inhibitors

doi: 10.1016/j.isci.2023.107302

Figure Lengend Snippet:

Article Snippet: Mouse Monoclonal EP-CAM (C10) Antibody , Santa Cruz , Cat# sc-25308: RRID: AB_62531.

Techniques: Control, Virus, Recombinant, Live Cell Imaging, Chromatin Immunoprecipitation, Luciferase, CCK-8 Assay, Small Interfering RNA, shRNA, Software

Journal: Anti-cancer drugs

Article Title: Bcl-2 and Bcl-xL mediate resistance to receptor tyrosine kinase targeted therapy in lung and gastric cancer

doi: 10.1097/CAD.0000000000000561

Figure Lengend Snippet: Induction of Bcl-2 and Bcl-xL by TKIs in RTK-dependent lung and gastric cancer cell lines. EGFR-dependent lung cancer cell lines HCC827, PC9, and HCC4006 (A), ALK-dependent lung cancer cell line H3122, and HER2-dependent gastric cancer cell line NCI-N87 were treated with gefitinib (1 μM), gefitinib (0.2 μM), erlotinib (1 μM), TAE684 (0.3 μM), lapatinib (0.1 μM) respectively for 6, 24, and 48 hrs. Expression of indicated proteins was assessed by immunoblots.

Article Snippet: The small molecule inhibitors TAE684 (ALK TKI), Lapatinib (Dual EHER2/EGFR TKI), and ABT-263 (Bcl-2 and Bcl-xL inhibitor) were obtained from Selleck Biochemicals.

Techniques: Expressing, Western Blot

Journal: Anti-cancer drugs

Article Title: Bcl-2 and Bcl-xL mediate resistance to receptor tyrosine kinase targeted therapy in lung and gastric cancer

doi: 10.1097/CAD.0000000000000561

Figure Lengend Snippet: Combination of Bcl-2/Bcl-xL inhibitor ABT-263 and a TKI is superior to the TKI alone. (A) Cell viability was determined after cells were treated as indicated for 72 hrs. Erlotinib, TAE684, Lapatinib, and ABT-263 were used at 1, 0.3, 0.1, and 1 μM respectively. Results shown are the mean ± SD. *p ˂0.02 compared with DMSO control. (B) Crystal violet staining of cells treated with DMSO or ABT-263 (1 μM) for 4 days, and TKI (1 μM erlotinib, 0.3 μM TAE684, 0.1 μM Lapatinib), or the combination of TKI and ABT-263 for 4 weeks. Representative results are shown.

Article Snippet: The small molecule inhibitors TAE684 (ALK TKI), Lapatinib (Dual EHER2/EGFR TKI), and ABT-263 (Bcl-2 and Bcl-xL inhibitor) were obtained from Selleck Biochemicals.

Techniques: Control, Staining

Journal: Anti-cancer drugs

Article Title: Bcl-2 and Bcl-xL mediate resistance to receptor tyrosine kinase targeted therapy in lung and gastric cancer

doi: 10.1097/CAD.0000000000000561

Figure Lengend Snippet: Combination of inhibitors of PI3K/AKT and MEK/ERK is effective in parental but not in resistant cells. Cells were treated with indicated inhibitors. Gefitinib, TAE684, Lapatinib, and PHA665752 were used at 1, 0.3, 0.1, and 1 μM respectively. BEZ235 was used at 0.5 μM and GSK1120212 was used at 1 μM. Expression of indicated proteins was assessed by immunoblots (left panels). Cell viability was determined after cells were treated for 72 hrs (right panels). Results shown are the mean ± SD.

Article Snippet: The small molecule inhibitors TAE684 (ALK TKI), Lapatinib (Dual EHER2/EGFR TKI), and ABT-263 (Bcl-2 and Bcl-xL inhibitor) were obtained from Selleck Biochemicals.

Techniques: Expressing, Western Blot

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: The expression of tumor CD73 was significantly associated with the risk of distant metastasis after surgical operation and poor survival outcome. A CD73 proteins were mainly expressed on the membrane of cancer cells. CD39 proteins were majorly expressed in the stromal cells and partially expressed in the inflammatory cells. B The H-score of tumor CD73 was statistically associated with the risk of distant metastasis (n = 421 and unpaired T-test p = 0.0353). C Stromal CD39 expression was not associated with the present of LVI, PNI and distant metastasis (n = 421) by unpaired T-test. D Tumor CD73 expression was significantly associated with pathological T stage (n = 421, p = 0.0008). Stromal CD39 expression was not associated with pathological T stage (n = 421 and p = 0.70). One-way ANOVA test. E Patients with high tumor CD73 expression were clinically associated with worsen disease-free survival (DFS) in CRC patients (n = 421, p = 0.0076). High stromal CD39 expression was associated with favorable DFS in CRC patients (n = 421 p = 0.0034). F Tumor CD73 expression was significantly associated with distant metastasis-free survival (DMFS) and DFS in stage II-III COAD patients who received adjuvant chemotherapy (n = 177, log-rank p = 0.0077). G Tumor CD73 expression was conversely associated with the density of CD8+ TILs and CD45.+ TILs within tumor microenvironment (TME). Unpaired t test (n = 421)

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Expressing, Membrane, Adjuvant

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Correlation between clinicopathologic parameters, DFS and OS

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques:

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Univariate and multivariate analysis of DFS and known prognostic factors in stage I-IV colon adenocarcinoma patients

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques:

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: CD73 attenuated OXP-induced anti-tumor immunity by converting ATP into adenosine. A The expression level of CD73 in five colorectal cancer cell lines. B HCT116 cells were infected with lentivirus carrying shRNA against CD73, then selected by puromycin (2 μg/mL) for 3 days. Cell lysates were analyzed by western blot. C HCT116shNC and HCT116shCD73 cells were treated with OXP for 6 h. The conditioned medium was harvested for ATP and AMP assay (n = 3). *p < 0.05. D HCT116 cells were treated with OXP (50 μM) and CD73-IN-1 (10 mM) for 6 h. The conditioned medium was harvested for ATP and AMP assay (n = 3). *p < 0.05 and ***p < 0.001. E The schematic diagram of co-culture experiment to evaluate the dendritic cells (DC) maturation and T cell activity. F OXP-treated HCT116shNC and HCT116shCD73 cells were co-cultured with THP1-immature dendritic cells (iDC) for 24 h. The level of DC maturation was analyzed by flow cytometry (n = 3). *p < 0.05 and **p < 0.01. G After coculture, Jurkat T cells were co-cultured with HCT116shNC/THP1-iDC and HCT116 shCD73/THP1-iDC for 15 h. The T cell activation was analyzed by flow cytometry (n = 3). *p < 0.05 and **p < 0.01. H Jurkat T cells were individually co-cultured with HCT116shNC and HCT116.shCD73 cells for 24 h. The cytotoxicity was analyzed by CCK assay (n = 3). *p < 0.05 and **p < 0.01

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Expressing, Infection, shRNA, Western Blot, Co-Culture Assay, Activity Assay, Cell Culture, Flow Cytometry, Activation Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Multivariate analysis of DFS and known prognostic factors in high-risk stage II-III carcinoma patients

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Adjuvant

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Genomic correlates of NT5E and CD8 signatures in the TCGA dataset A CD73/NT5E was highly expressed on tumor tissues compared to normal tissues. B High CD73 mRNA expression was remarkably associated with poor survival outcome in CRC patients that retrieved from TCGA database (cut-off mRNA = 9.0, n = 368, p = 0.0453). Moreover, CD39 mRNA expression was not associated with poor survival outcome in CRC patients that retrieved from TCGA database (cut-off mRNA = 3.37, n = 515, p = 0.7079). C High CD73 mRNA expression was conversely associated with less expression of T-cell mediated signatures (TCGA database, n = 368, unpaired T test). ***p < 0.001

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Blockade of CD73 enhanced the therapeutic efficacy of immunogenic chemotherapy. A The schematic diagram of treatment regimen. BALB/c mice were subcutaneous injected with 2 × 105 CT26 cells for 5 days and intravenous injected with 1 × 10.5 CT26 cells on Day 5 (n = 5). Oxaliplatin (OXP, 5 mg/kg, i.p.) was administrated on Day 7, 10, 13, 16 and 19. Anti-CD73 mAb (clone TY/23, 100 μg/mouse) and IgG control mAb were administrated on Day 8, 11, 14 and 17. B The tumor volume was measured every 2–3 days (n = 5). **p < 0.01 and ***p < 0.001. C The tumor volume on day 30 was shown (n = 5). *p < 0.05 and ***p < 0.001. D The resected tumors were weighed (n = 4). *p < 0.05 and ***p < 0.001. E The representative images of primary tumor and apoptotic marker cleaved caspase-3 (n = 4). *p < 0.05 and ***p < 0.001. F The representative images of lung were shown (n = 4). The number of lung metastatic nodules was evaluated under microscope. *p < 0.05, *p < 0.01 and ***p < 0.001. G The mouse survival was assessed with Kaplan–Meier survival curves (n = 6)

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Drug discovery, Injection, Control, Marker, Microscopy

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting CD73 increases therapeutic response to immunogenic chemotherapy by promoting dendritic cell maturation

doi: 10.1007/s00262-023-03416-4

Figure Lengend Snippet: Blockade of CD73 enhanced DC infiltration for anti-tumor immunity when combined with OXP treatment. A The resected tumors were stained with CD11 (DC marker) and GzmB (cytotoxic T cell marker), and evaluated under immunofluorescent microscope. B The quantification of CD11c+ DCs was shown (n = 4). *p < 0.05 and ***p < 0.001. C The quantification of GzmB+ T cells was shown (n = 4). *p < 0.05 and ***p < 0.001. D The tumor-infiltrating immune cells were extracted from resected tumors for flow cytometry. The absolute number of CD11c+MHC-II+ DCs was shown (n = 3–4). *p < 0.05 and ***p < 0.001 (n = 3). E The absolute number of tumor-infiltrating CD86+CD80+CD11c+MHC-II+ DCs was shown (n = 3–4). *p < 0.05 and ***p < 0.001. F The absolute number of tumor-infiltrating CD4+T, CD8+T and CD44+CD62L−CD8+TEM cells was shown (n = 3–4). *p < 0.05 and ***p < 0.001. G The absolute number of tumor-infiltrating cytotoxic GzmB+CD8+T cells was shown (n = 3–4). *p < 0.05 and ***p < 0.001. H The absolute number of tumor-infiltrating cytotoxic IFNγ+CD8.+T cells was shown (n = 3–4). *p < 0.05 and ***p < 0.001

Article Snippet: Small molecule CD73 inhibitor (CD73-IN-1, Cat. HY-103695) was purchased from MedChemExpress (CA, USA).

Techniques: Staining, Marker, Microscopy, Flow Cytometry