Journal: Cancer Research

Article Title: A Specific STAT3-Binding Peptide Exerts Antiproliferative Effects and Antitumor Activity by Inhibiting STAT3 Phosphorylation and Signaling

doi: 10.1158/0008-5472.can-13-2187

Figure Lengend Snippet: Figure 1. Characterization of APTSTAT3, a specific STAT3-binding peptide. A, schematic depiction of the structure of a STAT3-specific aptide (APTSTAT3) and control scrambled peptide (APTscr) with amino acid sequences and a cell permeable form of the STAT3-specific aptide (APTSTAT3-9R). B, phage ELISAs using a phage displaying APTSTAT3 sequence against various proteins (STAT3, streptavidin, BSA, VEGF, TNF-a, and anti-His6-tag-antibody; left). C, concentration–escalation ELISAs using APTSTAT3, APTSTAT3-9R, and APTscr-9R peptides against STAT3 protein. Antiaptide (trpzip) mAb was used to detect. D, confocal microscopy images of A549 human lung carcinoma cells after treatment with none (control), FITC-APTSTAT3-9R, and FITC-APTscr-9R.

Article Snippet: STAT3 protein was obtained from SignalChem.

Techniques: Binding Assay, Control, Sequencing, Concentration Assay, Confocal Microscopy

Journal: Cancer Research

Article Title: A Specific STAT3-Binding Peptide Exerts Antiproliferative Effects and Antitumor Activity by Inhibiting STAT3 Phosphorylation and Signaling

doi: 10.1158/0008-5472.can-13-2187

Figure Lengend Snippet: Figure 2. The inhibitory effects of APTSTAT3-9R on STAT3 signaling. A, DNA-binding assay for STAT3 in cell extracts of A549 human lung carcinoma following treatment with APTSTAT3-9R (7.5, 15, and 30 mmol/L) or APTscr-9R (30 mmol/L) for 6 hours. B, quantification of mRNA level of STAT3 downstream target genes, including Bcl-xL, cyclin D1, and survivin, in A549 cells following treatment with APTSTAT3-9R (7.5, 15, and 30 mmol/L) or APTscr-9R (30 mmol/L) for 6 hours. C, dose-dependent Western blot assays for STAT3 downstream target proteins (Bcl-xL, cyclin D1, and survivin) and P-STAT3 (Y705) in A549 cells following treatment with APTSTAT3-9R (7.5, 15, and 30 mmol/L) or APTscr-9R (30 mmol/L) for 6 hours. D, Western blot assays for STAT3, P-STAT3 (Y705), and other signaling in A549 cells following treatment with APTSTAT3-9R and APTscr-9R (30 mmol/L) for 6 hours. , P < 0.005 (one-way ANOVA); , P < 0.001.

Article Snippet: STAT3 protein was obtained from SignalChem.

Techniques: DNA Binding Assay, Western Blot

Journal: Cancer Research

Article Title: A Specific STAT3-Binding Peptide Exerts Antiproliferative Effects and Antitumor Activity by Inhibiting STAT3 Phosphorylation and Signaling

doi: 10.1158/0008-5472.can-13-2187

Figure Lengend Snippet: Figure 4. Antitumor activity of APTSTAT3-9R in an A549 xenograft model. A, the tumor growth study in the A549 xenograft model. Nude mice bearing A549 tumors (50 mm3) were treated four times with APTSTAT3-9R (8 mg/kg), APTscr-9R (8 mg/kg), or PBS. Tumor volumes were measured every other day using calipers. Mean tumor size SEM (n ¼ 5). Arrows indicate the injection day. B, a photograph of tumors excised from mice after 30 days of treatment. C, Western blot assays for STAT3 and P-STAT3 (Y705) in the excised tumor tissues. D, TUNEL assays of tumor sections. , P < 0.001 (one-way ANOVA).

Article Snippet: STAT3 protein was obtained from SignalChem.

Techniques: Activity Assay, Injection, Western Blot, TUNEL Assay

Journal: Journal of Diabetes Investigation

Article Title: Cordyceps cicadae polysaccharides attenuate diabetic nephropathy via the miR ‐30a‐3p/ TRIM16 axis

doi: 10.1111/jdi.14116

Figure Lengend Snippet: PCR primers used in this study

Article Snippet: The renal sections, after being deparaffinized and rehydrated, were incubated overnight at 4°C with TRIM16 primary antibodies (A09514‐4, 1:500; Boster Biological Engineering Co., International Enterprise Center, Wuhan, China), Bcal‐2 (ab182858, 1:500; Abcam), and bcl‐2‐associated X protein (BAX) (ab53154, 1:500; Abcam); and P‐cadherin antibodies (ab242060, 1:500; Abcam) and N‐cadherin (ab76011, 1:500; Abcam); then incubated with Alexa Fluor 594‐conjugated (red) goat anti‐mouse IgG and Alexa Fluor 488‐conjugated (green) goat anti‐rabbit IgG (1:1000; Invitrogen, Carlsbad, CA, USA) secondary antibodies at 25°C for 1 h. Nuclei were counterstained with a Hoechst solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques:

Journal: Journal of Diabetes Investigation

Article Title: Cordyceps cicadae polysaccharides attenuate diabetic nephropathy via the miR ‐30a‐3p/ TRIM16 axis

doi: 10.1111/jdi.14116

Figure Lengend Snippet: Cordyceps cicadae polysaccharides administration attenuated cell apoptosis and epithelial–mesenchymal transition in the kidneys of diabetic nephropathy mice. (a, b) At the end of 12 weeks of treatment, the expression of miR ‐30a‐3p and TRIM16 was determined in the kidney tissues of mice in each group by RT‐PCR , n = 8. (c) The protein expression level of TRIM16 in kidney tissues of mice was analyzed by western blot, n = 5. (d, e) Protein expression of TRIM16 in the kidney tissues using IHC staining, scale bar = 50 μm, n = 5. (f) TUNEL staining was used to detect cell apoptosis in the kidney tissues, scale bar, 20 μm, n = 5. (g) Protein expression of Bcl‐2 and Bax in the kidney tissues were detected by IHC staining, scale bar = 50 μm, n = 5. (h) Representative images of immunofluorescence staining for P‐cadherin (red), N‐cadherin (green), and DAPI (blue), scale bar, 50 μm, n = 5, scale bar = 20 μm. Data are expressed as mean ± SD . * P < 0.05, ** P < 0.01.

Article Snippet: The renal sections, after being deparaffinized and rehydrated, were incubated overnight at 4°C with TRIM16 primary antibodies (A09514‐4, 1:500; Boster Biological Engineering Co., International Enterprise Center, Wuhan, China), Bcal‐2 (ab182858, 1:500; Abcam), and bcl‐2‐associated X protein (BAX) (ab53154, 1:500; Abcam); and P‐cadherin antibodies (ab242060, 1:500; Abcam) and N‐cadherin (ab76011, 1:500; Abcam); then incubated with Alexa Fluor 594‐conjugated (red) goat anti‐mouse IgG and Alexa Fluor 488‐conjugated (green) goat anti‐rabbit IgG (1:1000; Invitrogen, Carlsbad, CA, USA) secondary antibodies at 25°C for 1 h. Nuclei were counterstained with a Hoechst solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunohistochemistry, TUNEL Assay, Staining, Immunofluorescence

Journal: Journal of Diabetes Investigation

Article Title: Cordyceps cicadae polysaccharides attenuate diabetic nephropathy via the miR ‐30a‐3p/ TRIM16 axis

doi: 10.1111/jdi.14116

Figure Lengend Snippet: Expression of miR‐30a‐3p and TRIM16 in MPC5 podocytes treated with high glucose. The MPC5 podocytes were treated with normal glucose, mannitol, and high glucose conditions for 6, 12, and 24 h. (a) The expression level of miR‐30a‐3p in mouse podocyte was detected by RT‐qPCR. (b, c) The mRNA and protein expression of TRIM16 was determined by RT‐PCR and western blotting. * P < 0.05, ** P < 0.01.

Article Snippet: The renal sections, after being deparaffinized and rehydrated, were incubated overnight at 4°C with TRIM16 primary antibodies (A09514‐4, 1:500; Boster Biological Engineering Co., International Enterprise Center, Wuhan, China), Bcal‐2 (ab182858, 1:500; Abcam), and bcl‐2‐associated X protein (BAX) (ab53154, 1:500; Abcam); and P‐cadherin antibodies (ab242060, 1:500; Abcam) and N‐cadherin (ab76011, 1:500; Abcam); then incubated with Alexa Fluor 594‐conjugated (red) goat anti‐mouse IgG and Alexa Fluor 488‐conjugated (green) goat anti‐rabbit IgG (1:1000; Invitrogen, Carlsbad, CA, USA) secondary antibodies at 25°C for 1 h. Nuclei were counterstained with a Hoechst solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Journal of Diabetes Investigation

Article Title: Cordyceps cicadae polysaccharides attenuate diabetic nephropathy via the miR ‐30a‐3p/ TRIM16 axis

doi: 10.1111/jdi.14116

Figure Lengend Snippet: miR‐30a‐3p regulates TRIM16 in MPC5 podocytes. (a) TRIM16 is a predicted target of miR‐30a‐3p by TargetScan. (b) A luciferase reporter plasmid containing wildtype or mutant TRIM16 was cotransfected into MPC5 podocytes with miR‐30a‐3p or miR‐NC. Luciferase activity was determined at 48 h after transfection using the dual‐luciferase assay and was normalized to Renilla activity. (c–e) RT‐PCR and western blot analysis of TRIM16 in podocytes transfected with miR‐30a‐3p or miR‐NC or miR‐30a‐3p inhibitor or inhibitor of negative control. Data are presented as mean ± SD, n = 3. ** P < 0.01, *** P < 0.001. ## P < 0.001.

Article Snippet: The renal sections, after being deparaffinized and rehydrated, were incubated overnight at 4°C with TRIM16 primary antibodies (A09514‐4, 1:500; Boster Biological Engineering Co., International Enterprise Center, Wuhan, China), Bcal‐2 (ab182858, 1:500; Abcam), and bcl‐2‐associated X protein (BAX) (ab53154, 1:500; Abcam); and P‐cadherin antibodies (ab242060, 1:500; Abcam) and N‐cadherin (ab76011, 1:500; Abcam); then incubated with Alexa Fluor 594‐conjugated (red) goat anti‐mouse IgG and Alexa Fluor 488‐conjugated (green) goat anti‐rabbit IgG (1:1000; Invitrogen, Carlsbad, CA, USA) secondary antibodies at 25°C for 1 h. Nuclei were counterstained with a Hoechst solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Luciferase, Plasmid Preparation, Mutagenesis, Activity Assay, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Negative Control

Journal: Journal of Diabetes Investigation

Article Title: Cordyceps cicadae polysaccharides attenuate diabetic nephropathy via the miR ‐30a‐3p/ TRIM16 axis

doi: 10.1111/jdi.14116

Figure Lengend Snippet: Cordyceps cicadae polysaccharides protect mouse podocyte from high glucose induced apoptosis and epithelial–mesenchymal transition by controlling miR‐30a‐3p expression. (a) Relative miR‐30a‐3p and TRIM16 expression were detected by RT‐PCR and western blotting analysis. (b) Cell viability was determined by MTT assay. (c, d) The apoptosis of cells as detected by flow cytometry. (e) Representative images of TUNEL staining to illustrate apoptotic MPC5 podocytes in each group, cell nuclei were labeled in blue by DAPI, scale bar, 50 μm. (f) Representative immunofluorescence images of E‐cadherin (E‐cad, red) and N‐cadherin (N‐cad, green), cell nuclei were labeled in blue by DAPI, scale bar, 20 μm. Data are presented as mean ± SD, n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The renal sections, after being deparaffinized and rehydrated, were incubated overnight at 4°C with TRIM16 primary antibodies (A09514‐4, 1:500; Boster Biological Engineering Co., International Enterprise Center, Wuhan, China), Bcal‐2 (ab182858, 1:500; Abcam), and bcl‐2‐associated X protein (BAX) (ab53154, 1:500; Abcam); and P‐cadherin antibodies (ab242060, 1:500; Abcam) and N‐cadherin (ab76011, 1:500; Abcam); then incubated with Alexa Fluor 594‐conjugated (red) goat anti‐mouse IgG and Alexa Fluor 488‐conjugated (green) goat anti‐rabbit IgG (1:1000; Invitrogen, Carlsbad, CA, USA) secondary antibodies at 25°C for 1 h. Nuclei were counterstained with a Hoechst solution (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, MTT Assay, Flow Cytometry, TUNEL Assay, Staining, Labeling, Immunofluorescence

Journal: RSC Advances

Article Title: p-JAK2 plays a key role in catalpol-induced protection against rat intestinal ischemia/reperfusion injury

doi: 10.1039/c7ra10506a

Figure Lengend Snippet: Fig. 4 Catalpol inhibited apoptosis via targeting JAK2/STAT3 signaling pathway in I/R-injured rats. (A) TUNEL staining of intestine after intestinal I/R. (B) Representative western blot analysis of total and phosphorylated JAK2 and total and phosphorylated STAT3 (n ¼ 3). (C) The protein expression of Bcl-2, Bax and cleaved caspase3 in intestine (n ¼ 3). Values are expressed as mean SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. sham group; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. I/R group.

Article Snippet: Rabbit polyclonal antibodies were used as primary antibodies against p-JAK2, JAK2, p-STAT3, STAT3 (Abclonal, Woburn, USA), Bcl-2, Bax, cleaved caspase3, occludin (Proteintech, Wuhan, China), and ZO-1 (Santa Cruz, CA, USA).

Techniques: TUNEL Assay, Staining, Western Blot, Expressing

Journal: RSC Advances

Article Title: p-JAK2 plays a key role in catalpol-induced protection against rat intestinal ischemia/reperfusion injury

doi: 10.1039/c7ra10506a

Figure Lengend Snippet: Fig. 6 Catalpol inhibited JAK2/STAT3 signaling pathway in H/R-injured IEC-6 cells. (A) Effects of catalpol on the expression of p-JAK2, JAK2, p- STAT3, STAT3 in IEC-6 cells in normal and H/R conditions (n ¼ 3). (B) Effects of catalpol on the expression of Bcl-2, Bax and cleaved caspase3 in IEC-6 cells in normal and H/R conditions (n ¼ 3). (C) Effects of catalpol on p-STAT3 translocation in IEC-6 cells in normal and H/R conditions (original magnification 400). Values are expressed as mean SEM. **P < 0.01, ***P < 0.001 vs. control group; #P < 0.05, ##P < 0.01 vs. H/R group.

Article Snippet: Rabbit polyclonal antibodies were used as primary antibodies against p-JAK2, JAK2, p-STAT3, STAT3 (Abclonal, Woburn, USA), Bcl-2, Bax, cleaved caspase3, occludin (Proteintech, Wuhan, China), and ZO-1 (Santa Cruz, CA, USA).

Techniques: Expressing, Translocation Assay, Control

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: The role and mechanism of TLR4-siRNA in the impairment of learning and memory in young mice induced by isoflurane.

doi: 10.17219/acem/147047

Figure Lengend Snippet: Fig. 6. Brain-derived neurotrophic factor (BDNF) and cyclic adenosine monophosphate response element-binding protein 1 (CREB1) mRNA expressions in the hippocampal tissue of each group detected with quantitative real-time polymerase chain reaction (qRT-PCR). Tukey’s honestly significant difference (HSD); compared with the control group, *p < 0.05; compared with the isoflurane group, #p < 0.05 (n = 3/group). The interval represents the mean value. The ends of the interval represent standard deviation (SD)

Article Snippet: The reagents used in the study included: isoflurane (national drug approval No. H20070172; Shanghai Hengrui Pharmaceutical Co., Ltd., Shanghai, China); TLR4-siRNA oligos set (cat. No. i548002; Applied Biological Materials Inc., Richmond, Canada); TRIzon reagent (CW0580S), ultrapure RNA extraction kit (CW0581M), HiFiScript first strand complementary DNA (cDNA) synthesis kit (CW2569M), UltraSYBR Mixture (CW0957M), bicinchoninic acid (BCA) protein assay kit (CW0014S) (Beijing ComWin Biotech Co., Ltd., Beijing, China); polyvinylidene difluoride (PVDF) membrane (IPVH00010; EMD Millipore Corporation, Billerica, USA); marker (PageRulerTM Prestained Protein Ladder; #26617), super enhanced chemiluminescence (ECL) plus (RJ239676; Thermo Fisher Scientific, Waltham, USA); mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1/2000, TA-08), goat anti-rabbit immunoglobulin G (IgG) (H+L) horseradish peroxidase (HRP) conjugate secondary antibody (1/5000, ZB-2301; ZSGB-BIO, Beijing, China); rabbit anti-TLR4 polyclonal antibody (1/1000, bs-20594R), rabbit anti-interleukin (IL)-6 polyclonal antibody (1/500, bs6309R; Bioss Antibodies Inc., Woburn, USA); rabbit antitumor necrosis factor alpha (TNF-α) polyclonal antibody (1/500, AF7014; Affinity Biosciences, Cincinnati, USA); rabbit anti-brain-derived neurotrophic factor (BDNF) (1/1000, ab108319; Abcam, Cambridge, USA); rabbit anti-cyclic adenosine monophosphate response element-binding protein 1 (CREB1, 1/1000, 12208-1-ap), rabbit anti-extracellular signal-regulated kinase 1/2 (ERK1/2, 1/1000, 16443-1-ap; Proteintech Group, Inc., Rosemont, USA); rabbit anti-c-Jun N-terminal kinase (JNK, 1/1000, df6089; Affinity Biosciences); terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay kit (C1088; Beyotime Biotechnology, Shanghai, China); mouse TNF-α enzyme-linked immunosorbent assay (ELISA) kit (MM-0132M1), mouse IL-6 kit (MM-0163M1; Jiangsu Meimian Industrial Co., Ltd., Jiangsu, China); radioimmunoprecipitation assay (RIPA) lysis buffer (C1053), skimmed milk powder (P1622; Applygen Technologies Inc., Beijing, China); and bovine serum albumin (BSA, A8020; Solarbio Life Sciences, Beijing, China).

Techniques: Derivative Assay, Binding Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: β-Glucan produced by Lentinus edodes suppresses breast cancer progression via the inhibition of macrophage M2 polarization by integrating autophagy and inflammatory signals.

doi: 10.1002/iid3.876

Figure Lengend Snippet: FIGURE 1 Expression of Nur77 and p62 in human breast cancer tissue samples. (A) Expression of Nur77, p62, CD68, and Ki‐67 proteins in T and NT tissue samples as per the immunohistochemical analysis. Scale bar: 50 μm. (B) mRNA levels of Nur77 and p62 in T and NT tissue samples as per qPCR. (C) Expression of Nur77 and p62 proteins in T and NT tissue samples as per western blot analysis analysis (n = 3). Data are shown as mean ± SD, n ≥3. *p < .05; **p < .01. NT, nontumor; qPCR, quantitative real‐time polymerase chain reaction; T, tumor.

Article Snippet: Next, nonspecific protein binding was blocked by incubation the sections with 2% bovine serum albumin and with primary antibody against Nur77 (12235AD; Proteintech), p62 (AF5384; Affinity), CD68 (ab955; Abcam), and Ki67 (ab15580; Abcam) overnight at 4°C; then, the secondary antibody was added, carried with horseradish peroxidase, and developed according to the manufacturer's protocol (IHC staining module; Beijing Zhongshan Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Western Blot, Real-time Polymerase Chain Reaction

Journal: Immunity, inflammation and disease

Article Title: β-Glucan produced by Lentinus edodes suppresses breast cancer progression via the inhibition of macrophage M2 polarization by integrating autophagy and inflammatory signals.

doi: 10.1002/iid3.876

Figure Lengend Snippet: FIGURE 3 Effect of Lentinus edodes (LNT) on cell proliferation and cell apoptosis in tumor tissues and breast cancer cell lines. (A) Hematoxylin and eosin (H&E) staining (the top images) was used to investigate inflammation and infiltration in the LNT‐treated group and the control group and for the examination of lung metastasis (lower images) of LNT‐treated group and control group. (B) Immunofluorescence staining was used to detect the expression of proliferation and apoptosis‐associated proteins using antibodies against PCNA in the LNT‐treated and control groups (upper images), and TUNEL assay was performed to assess cell apoptosis in the tumor tissue of LNT‐treated and control groups (lower images). (C) Expression of Nur77, p62, LC3, p‐mTOR, p‐AKT, p‐IKK, IKBα, PCNA, cleaved caspase‐3, and GAPDH proteins in tumor tissues as per western blot analysis analysis. (D) Expression of Nur77, p62, LC3, p‐mTOR, p‐AKT, p‐IKK, IKBα, PARP1, and β‐actin proteins in tumor cells as per western blot analysis analysis. Data are shown as mean ± SD, n ≥3.

Article Snippet: Next, nonspecific protein binding was blocked by incubation the sections with 2% bovine serum albumin and with primary antibody against Nur77 (12235AD; Proteintech), p62 (AF5384; Affinity), CD68 (ab955; Abcam), and Ki67 (ab15580; Abcam) overnight at 4°C; then, the secondary antibody was added, carried with horseradish peroxidase, and developed according to the manufacturer's protocol (IHC staining module; Beijing Zhongshan Biotechnology).

Techniques: Staining, Control, Immunofluorescence, Expressing, TUNEL Assay, Western Blot

Journal: Immunity, inflammation and disease

Article Title: β-Glucan produced by Lentinus edodes suppresses breast cancer progression via the inhibition of macrophage M2 polarization by integrating autophagy and inflammatory signals.

doi: 10.1002/iid3.876

Figure Lengend Snippet: FIGURE 4 Macrophage polarization in tumor tissues and tumor cells. (A) Immunofluorescence analysis was used to detect the number of CD206‐ and CD86‐positive cells in the Lentinus edodes (LNT)‐treated and control groups; CD206‐ (red) and CD86‐positive (green) cells indicate M2 and M1 macrophages, respectively. (B) Schematic diagram of cell supernatant transfer culture. (C) T47D cells were treated with LNT, and the expression of Nur77, p62, LC3, PARP1, and β‐actin proteins was analyzed in different groups using western blot analysis. (D) Macrophages treated with culture medium from LNT‐treated T47D cells was used to treat macrophages, and the expression of iNOS, Arg1, and β‐actin proteins in different groups was analyzed using western blot analysis. (E) T47D cells treated with culture medium from macrophages (pretreated with medium of LNT‐treated T47D cells); expression of Nur77, p62, LC3, PARP1, and β‐actin proteins in different groups as per western blot analysis analysis. (F) Schematic diagram of macrophage‐directed differentiation culture induced by cytokines with different properties. (G) Expression of iNOS, Arg1, and β‐actin proteins in different groups as per western blot analysis analysis. (H) Semiquantitative analysis of iNOS and Arg1 proteins in panel (G). Data are shown as mean ± SD, n ≥3. *p < .05; **p < .01; ***p < .001; ****p < .0001.

Article Snippet: Next, nonspecific protein binding was blocked by incubation the sections with 2% bovine serum albumin and with primary antibody against Nur77 (12235AD; Proteintech), p62 (AF5384; Affinity), CD68 (ab955; Abcam), and Ki67 (ab15580; Abcam) overnight at 4°C; then, the secondary antibody was added, carried with horseradish peroxidase, and developed according to the manufacturer's protocol (IHC staining module; Beijing Zhongshan Biotechnology).

Techniques: Immunofluorescence, Control, Expressing, Western Blot

Journal: Immunity, inflammation and disease

Article Title: β-Glucan produced by Lentinus edodes suppresses breast cancer progression via the inhibition of macrophage M2 polarization by integrating autophagy and inflammatory signals.

doi: 10.1002/iid3.876

Figure Lengend Snippet: FIGURE 5 Effects of LNT on TNF‐α‐mediated autophagic cell death in breast cancer cells. T47D cells were transfected with Nur77 siRNA and untransfected cells were used as control. The cells were treated with 20 ng/mL TNF‐α alone or a combination of TNF‐α and 1000 μg/mL LNT. (A) Expression of Nur77, p62, LC3, PCNA, caspase‐3, and β‐actin proteins in different groups as per western blot analysis analysis. (B) Expression of pIKBα, IKBα, p‐IKK, p‐mTOR, p‐AKT, and GAPDH proteins in different groups as per western blot analysis analysis. Data are shown as mean ± SD, n ≥3. LNT, Lentinus edodes; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Next, nonspecific protein binding was blocked by incubation the sections with 2% bovine serum albumin and with primary antibody against Nur77 (12235AD; Proteintech), p62 (AF5384; Affinity), CD68 (ab955; Abcam), and Ki67 (ab15580; Abcam) overnight at 4°C; then, the secondary antibody was added, carried with horseradish peroxidase, and developed according to the manufacturer's protocol (IHC staining module; Beijing Zhongshan Biotechnology).

Techniques: Transfection, Control, Expressing, Western Blot

Journal: Immunity, inflammation and disease

Article Title: β-Glucan produced by Lentinus edodes suppresses breast cancer progression via the inhibition of macrophage M2 polarization by integrating autophagy and inflammatory signals.

doi: 10.1002/iid3.876

Figure Lengend Snippet: FIGURE 6 Schematic diagram of Lentinus edodes (LNT)‐mediated improvement in the progression of breast cancer. LNT promoted the secretion of cytokines via the regulation of Nur77 and the downstream mTOR/AKT and p62/LC3‐mediated autophagy. The cytokines subsequently inhibited macrophage M2 polarization, which inhibited the AKT/mTOR and NF‐κB signaling axes in breast cancer cells, thereby promoting autophagic tumor cell death and inhibiting breast cancer progression.

Article Snippet: Next, nonspecific protein binding was blocked by incubation the sections with 2% bovine serum albumin and with primary antibody against Nur77 (12235AD; Proteintech), p62 (AF5384; Affinity), CD68 (ab955; Abcam), and Ki67 (ab15580; Abcam) overnight at 4°C; then, the secondary antibody was added, carried with horseradish peroxidase, and developed according to the manufacturer's protocol (IHC staining module; Beijing Zhongshan Biotechnology).

Techniques:

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) IER3IP1 protein levels in IER3IP1 KD MIN6 cells. MIN6 cells were infected with a lentivirus vector that drives expression of shRNA that targets the IER3IP1 transcript (IER3IP1 KD) or control lentivirus vector that does not have a specific target. IER3IP1 protein levels were measured 4 days after infection by western blot (n=3). The loaded proteins in the two “- or +” lines were the same but from the different plates. (B) Cell death was determined by PI-staining in MIN6 cells after infection with IER3IP1 shRNA lentivirus. (C) Cleaved caspase3 protein levels in IER3IP1 KD cells. 4 days after infection with IER3IP1 shRNA lentivirus, cleaved caspase3 and PARP proteins were assayed by western blot (n=3). The loaded proteins in the two “- or +” lines were the same but from the different plates. (D) Z-VAD inhibits caspase3 cleavage induced by IER3IP1 KD. MIN6 cells were treated with the caspase inhibitor Z-VAD (20 μM), for 2 hours prior to infection with IER3IP1 shRNA lentivirus. 4 days later, cleaved caspase3 protein was assayed by western blot (n=3). (E) Z-VAD inhibits cell death induced by IER3IP1 KD in MIN6 cells. MIN6 cells were treated with 20 μM Z-VAD, for 2 hours prior to infection with IER3IP1 shRNA lentivirus and cell death was determined by PI-staining on day 4 (n=3). ***P<0.001 compared to control group. ###P<0.001 compared to IER3IP1 KD group. (F) TUNEL labeling of IER3IP1 KD MIN6 cells. 4 days after infection with IER3IP1 shRNA lentivirus, apoptotic cells were assayed by TUNEL staining. Quantitative TUNEL data are shown. Scale bar, 20 μm. ***P<0.001 compared to control group. Values are mean ± SEM.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Infection, Plasmid Preparation, Expressing, shRNA, Control, Western Blot, Staining, TUNEL Assay, Labeling

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A+B) MIN6 cells were treated with IER3IP1 shRNA lentivirus for 3 days. Cells were then stained with TMRE dye to measure ΔΨ m . Scale bar, 20 μm. ***P<0.001. (C+D) IER3IP1 suppression induces cytochrome c release. 3 days after IER3IP1 KD in MIN6 cells, the images were taken under fluorescence microscopy (C). The green, red and blue staining represent cytochrome c, HSP60 and DAPI, respectively. Scale bar, 20 μm. The release of cytochrome c was also measured (D). **P<0.01 compared to control group. Values are mean ± SEM.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: shRNA, Staining, Fluorescence, Microscopy, Control

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) Bax and Bak protein levels in control and IER3IP1 KD cells. 3 days after IER3IP1 KD in MIN6 cells, Bax and Bak protein levels showed no difference between control and IER3IP1 KD cells (n=3). The loaded proteins in the two “- or +” lines were the same but from the different plates. (B) Immunoprecipitate of Bax. 3 days after IER3IP1 KD in MIN6 cells, cells were lysed in 1% CHAPS and then immunoprecipitated with the 6A7 anti-BAX antibody. Immunoprecipitates were analyzed by anti-BAX (N20) immunoblots. (C) Bax KD inhibits IER3IP1 KD-induced Bax activation. 3 days after IER3IP1 KD in MIN6 cells, the images were taken under fluorescence microscopy. Scale bar, 20 μm. (D) Bax KD inhibits cell death induced by IER3IP1 suppression. MIN6 cells were infected with IER3IP1 shRNA or IER3IP1/Bax double shRNA lentivirus. 4 days later, cell death was determined by PI-staining (n=3). ***P<0.001 compared to control group. ###P<0.001 compared to IER3IP1 KD group. Values are mean ± SEM.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Control, Immunoprecipitation, Western Blot, Activation Assay, Fluorescence, Microscopy, Infection, shRNA, Staining

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) Bim mRNA levels in control and IER3IP1 KD cells. 4 days after IER3IP1 KD in MIN6 cells, Bim mRNA levels were measured by real time quantitative reverse transcription-PCR (qRT-PCR) in MIN6 cells (n=3). ***P<0.001 compared to control group. Values are mean ± SEM. (B) Bim and Puma protein levels in IER3IP1 KD cells. 4 days after IER3IP1 KD in MIN6 cells, immunoblot analysis was performed to determine Bim, Puma and cleaved caspase3 protein levels in IER3IP1 KD MIN6 cells. The bar graph depicts the relative changes in the levels of the indicated proteins using densitometry analysis of the Western blots (n=3). Values are mean ± SEM. The loaded proteins in the two “- or +” lines were the same but from the different plates. (C) Western blot of IER3IP1/Bim DKD cells. 4 days after IER3IP1/Bim DKD in MIN6 cells, immunoblot of Bim and cleaved caspase3 in cells. (D) Measurement of cell death. 4 days after Bim/IER3IP1 DKD in MIN6 cells, cell death was determined by PI-staining (n=3). ***P<0.001 compared to control group. ###P<0.001 compared to IER3IP1 KD group. Values are mean ± SEM. (E+F) Western blot of IER3IP1 KD MIN6 cells. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of FoxO1/FoxO3a, FoxO1, FoxO3a (E) , p-Erk and Erk (F) were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Control, Reverse Transcription, Quantitative RT-PCR, Western Blot, Staining, Phospho-proteomics

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) Protein levels of anti-apoptotic Bcl-2 family members in IER3IP1 KD cells. 4 days after infection with IER3IP1 shRNA lentivirus, protein levels of Bcl-2, Bcl-xL and Mcl-1 in MIN6 cells were analyzed by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (B) Effect of overexpression of Bcl-xL on the cleavage of caspase3. MIN6 cells were infected with IER3IP1 shRNA lentivirus and retrovirus overexpressing Bcl-xL for 3 days, then protein levels of cleaved caspase3 and Bcl-xL were determined by Western blot. (C) Cell death was determined by PI-staining. MIN6 cells were infected with IER3IP1 shRNA lentivirus and Bcl-xL-overexpressed retrovirus, cell death was determined by PI-staining on day 4 and day 6 (n=3). ***P<0.001 compared to control group. ###P<0.001 compared to IER3IP1 KD group. Values are mean ± SEM.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Infection, shRNA, Western Blot, Over Expression, Staining, Control

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) mRNA levels of IRE1α, Perk and Atf6 in IER3IP1 KD cells. 4 days after infection with IER3IP1 shRNA lentivirus, mRNA levels of IRE1α, Perk and Atf6 in IER3IP1 KD cells were determined by qRT-PCR. *P<0.05, **P<0.01 compared to control group. Values are mean ± SEM. (B) Effect of IER3IP1 suppression on the IRE1α arm of the UPR. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of IRE1α, Bip, sXBP-1 and uXBP-were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (C) mRNA levels of downstream targets of sXBP-1 in IER3IP1 KD cells. 4 days after IER3IP1 KD in MIN6 cells, mRNA levels of downstream targets of sXBP-1, Erdj4, p58IPK, EDEM and Sec61a, were determined by qRT-PCR. Values are mean ± SEM. (D) Effect of IER3IP1 suppression on the Perk arm of the UPR. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of Perk and eIF2a, and ATF4 were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (E) IER3IP1 KD decreases UPR activation induced by ER stress in MIN6. 4 days after IER3IP1 KD, MIN6 cells were treated with 10 μg/ml tunicamycin (TC) or 1 μM thapsigargin (TG) for 6 hours. Then the expression levels of Chop, sXBP-1, uXBP-1 and IER3IP1 were determined by Western blot.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Infection, shRNA, Quantitative RT-PCR, Control, Phospho-proteomics, Western Blot, Activation Assay, Expressing

Journal: Oncotarget

Article Title: IER3IP1 deficiency leads to increased β-cell death and decreased β-cell proliferation

doi: 10.18632/oncotarget.18179

Figure Lengend Snippet: (A) Ki-67 staining of MIN6 cells. 2 days after IER3IP1 KD in MIN6 cells, Ki-67 staining for proliferation cell nuclei was performed in MIN6 cells. The Ki-67 staining was shown in the left. Scale bar, 20 μm. Quantitative Ki-67 staining data were shown in the right. Values are means ± SEM, *P < 0.05 compared to control cells. (B+C) Protein levels of phosphorylation of AKT, Erk, 4E-BP and S6. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of AKT (B) , 4E-BP and S6 (C) were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (D) Model presents how IER3IP1 suppression affects MIN6 cells. IER3IP1 suppression induces an increase in cell death and a decrease cell proliferation in MIN6 cells.

Article Snippet: Applied Biosystems (Foster City, CA) TaqMan assay numbers were: Mouse actin B, 4352933; IER3IP1, Mm01263577_g1; Bim, Mm00437796_m1 and Puma, Mm00519268_m1.

Techniques: Staining, Control, Phospho-proteomics, Western Blot

Journal: Nature Communications

Article Title: Therapeutic role of miR-19a/19b in cardiac regeneration and protection from myocardial infarction

doi: 10.1038/s41467-019-09530-1

Figure Lengend Snippet: miR-19a/19b reduces myocardial infarction-induced inflammation and cell death. a TUNEL staining (red) on transverse sections of adult hearts injected with miR-19a/19b or control mimic 4 days after MI. cTNT marks cardiomyocytes (green) and DAPI marks nuclei (blue). Scale bars = 48 μm. b , c Quantification of TUNEL-positive cardiomyocytes ( b ) and non-cardiomyocytes ( c ). n = 4 hearts, 6–14 fields per heart. d qRT-PCR detection of apoptosis gene expression. n = 3–7 hearts. e Western blot analysis of protein levels of PTEN, BIM, cleaved Caspase 3, and BAX in adult hearts. f Quantification of Western blot band density. n = 4 hearts. g Hierarchical clustering of 544 differentially expressed genes between miR-19a/19b and control mimic injected hearts 4 days after MI. Red and blue colors indicate up-regulated or down-regulated genes. h Gene ontology (GO) analysis of 393 down-regulated genes. ( i ) qRT-PCR analysis of expression of down-regulated genes related to immune response, cell death, and fibrosis. Up-regulated genes were also examined. NPPA serves as a control for cardiomyopathy. n = 3 hearts. j–l Immunohistochemistry analysis of M1 markers, ( j ) CD80, ( k ) iNOS, and M2 marker ( l ) Arg-1 in adult hearts injected with miR-19a/19b or control mimics at 48 and 72 h post MI. n = 4–8 hearts. Scale bars= 50 μm. m qRT-PCR analysis of expression of M1 and M2 marker genes and potential target genes in adult hearts. n = 3–6 hearts. n Western blot analysis of protein levels of SOCS and STAT3 pathways in adult hearts. o Quantification of Western blot band density. n = 3–4 hearts. Statistical significance was calculated using Student’s t -test in b , c , d , f , i , m , and o and data are presented as means ± s.e.m. * p < 0.05, ** p < 0.01 vs. control. Source data are provided as a Source Data file

Article Snippet: STAT3 protein was probed with mouse antibody to STAT3 (CST, #9139; 1:1000 dilution).

Techniques: TUNEL Assay, Staining, Injection, Control, Quantitative RT-PCR, Gene Expression, Western Blot, Expressing, Immunohistochemistry, Marker

Journal: Nature Communications

Article Title: Therapeutic role of miR-19a/19b in cardiac regeneration and protection from myocardial infarction

doi: 10.1038/s41467-019-09530-1

Figure Lengend Snippet: Therapeutic potential of miR-19a/19b in treating infarcted hearts. a Schematic of tail-vein injection of RNALancerII-delivered miRNA mimics post myocardial infarction (MI). b qRT-PCR detection of miR-19a and miR-19b expression in the heart 6 h after injection post MI injury. n = 1 heart each group. c qRT-PCR detection of miR-19a and miR-19b expression in the heart at 24 h after injection with or without MI injury. n = 2 hearts each group. d , e Echocardiography analyses of cardiac function after tail-vein injection of miR-19a/19b and control mimic at different time points post MI. d FS%, left ventricular fractional shortening. e LVIDs LV internal dimension at end-systole. n = 3–10 mice. f Representative images of series of transverse heart sections after tail-vein injection of miR-19a/19b or control mimics post MI. Sirius red/Fast green collagen staining marks myocardium (green) and scar (red). g Quantification of the size of scar. n = 3 hearts. h Western blot analysis of protein levels of SOCS and STAT3 in the heart 24 h after tail-vein injection of miR-19a/19b or control mimics post MI. Statistical significance was calculated using Student’s t -test in d , f , and g and data are presented as means ± s.e.m. * p < 0.05, ** p < 0.01 vs. control. Source data are provided as a Source Data file

Article Snippet: STAT3 protein was probed with mouse antibody to STAT3 (CST, #9139; 1:1000 dilution).

Techniques: Injection, Quantitative RT-PCR, Expressing, Control, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity

doi: 10.1038/s41419-021-03827-0

Figure Lengend Snippet: A Cell viability analysis after sorafenib treatment (24 h, 10 µM) measured by AnnexinV (left) and crystal violet (right). Characterization of sorafenib effect (24 h, 10 µM) by ( B ) mitochondrial ROS production (mitoSOX) and ( C ) gene expression analysis. Determination of mitochondrial function by ( D ) ROS production (mitoSOX) and ( E ) mitochondrial membrane potential (TMRE) at indicated short times of sorafenib treatment. * p < 0.05; ** p < 0.01; *** p < 0.001. (*miR-Ctrl vs. anti-miR-518d-5p; # compares sorafenib vs. non-treated ctrl cells. Data presented as mean ± SEM).

Article Snippet: The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 μM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium.

Techniques: Gene Expression, Membrane

Journal: Cell Death & Disease

Article Title: Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity

doi: 10.1038/s41419-021-03827-0

Figure Lengend Snippet: WB and qPCR analysis of c-Jun and PUMA in ( A ) BCLC3 and ( B ) Huh7 hepatoma cells under sorafenib treatment (24 h, 10 µM) and miR-518d inhibition/overexpression. Analysis of PUMA mitochondrial subcellular localization by double PUMA/MitoTracker staining under 3 h of sorafenib treatment in BCLC3 ( C ) and ( D ) Huh7 cells during miR-518d-5p inhibition/overexpression. Luciferase reporter assay of c-Jun-3′UTR in ( E ) BCLC3 and ( F ) Huh7 hepatoma cell lines after miR-518d-5p inhibition and overexpression, respectively. Normalized data referred to untreated control in each cell line. * p < 0.05; ** p < 0.01; *** p < 0.001. (*miR-Ctrl vs. miR-518d-5p; # compares sorafenib vs. non-treated ctrl cells. Data are represented as fold change referred to the non-treated control condition. Data presented as mean ± SEM).

Article Snippet: The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 μM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium.

Techniques: Inhibition, Over Expression, Staining, Luciferase, Reporter Assay, Control

Journal: Cell Death & Disease

Article Title: Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity

doi: 10.1038/s41419-021-03827-0

Figure Lengend Snippet: A Cell viability analysis after sorafenib treatment (24 h, 10 µM) measured by AnnexinV (left) and crystal violet (right). Sorafenib effect (24 h, 10 µM) by ( B ) mitochondrial ROS production (mitoSOX) and ( C ) gene expression analysis. Determination of mitochondrial function by analysis of ( D ) ROS production (mitoSOX) and ( E ) mitochondrial membrane potential (TMRE) at 3 h of sorafenib treatment. p < 0.05.; * p < 0.01; ** p < 0.001. (*miR-Ctrl vs. mimic-miR-518d-5p; # compares sorafenib vs. non-treated ctrl cells. Data are represented as fold change referred to the non-treated control condition. Data presented as mean ± SEM).

Article Snippet: The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 μM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium.

Techniques: Gene Expression, Membrane, Control

Journal: Cell Death & Disease

Article Title: Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity

doi: 10.1038/s41419-021-03827-0

Figure Lengend Snippet: A WB and qPCR analysis of c-Jun and PUMA in BCLC3 after sorafenib treatment (24 h, 10 µM) and miR-518d-5p inhibition and silencing of c-Jun. B Cell death analysis measured by AnnexinV and crystal violet and ( C ) qPCR analysis of indicated survival-related genes in response to miR-518d-5p inhibition and c-Jun silencing in sorafenib treated cells (24 h, 10 µM). Determination of ( D ) mitochondrial ROS production and ( E ) mitochondrial membrane potential after sorafenib treatment at indicated time points. F Analysis of PUMA mitochondrial subcellular localization by double PUMA/MitoTracker staining in sorafenib-treated cells (3 h, 10 µM). G Summary of the proposed model of miR-518d-5p role in sorafenib resistance in HCC through inhibition of c-Jun-mediated mitochondrial dysfunction and apoptosis. * p < 0.05; ** p < 0.01; *** p < 0.001.# compares sorafenib vs. non-treated ctrl cells. Data are represented as fold change referred to the non-treated control condition. Data presented as mean ± SEM.

Article Snippet: The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 μM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium.

Techniques: Inhibition, Membrane, Staining, Control

Journal: Cell Death & Disease

Article Title: Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity

doi: 10.1038/s41419-021-03827-0

Figure Lengend Snippet: A Time scheme of xenograft model. B Tumor volume fold increment in miR-Ctrl/miR-518d-5p xenograft tumors. C H&E, TUNEL, pan-RAS, and PCNA staining in tumor sections of xenografts tumors. D WB analysis with indicated antibody and ( E ) mRNA expression analysis of proteins and genes implicated in sorafenib antiproliferative and proapoptotic effect in murine xenografts. * p < 0.05; ** p < 0.01. Data are represented as fold change referred to as control. Data presented as mean ± SEM.

Article Snippet: The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 μM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium.

Techniques: TUNEL Assay, Staining, Expressing, Control