Journal: International Journal of Molecular Sciences

Article Title: Deletion of KLF10 Leads to Stress-Induced Liver Fibrosis upon High Sucrose Feeding

doi: 10.3390/ijms22010331

Figure Lengend Snippet: High sucrose diet activates stress-mediated inflammation and cell death in Klf 10 KO mouse livers. Eight-week-old WT and Klf 10 KO mice were fed either CD or HSD for eight weeks ( n = 5–6 mice/group). ( A ) Relative mRNA levels of proinflammatory cytokines Tnf α and Mcp 1 in the liver were analyzed via qPCR. * p < 0.05 vs. genotype-matched, CD-fed group and # p < 0.05 vs. diet-matched, genotype control. ( B ) Western blot analysis of p-JNK, JNK, p-ERK1/2, and ERK1/2, in the livers of WT and Klf 10 KO mice fed either CD or HSD. ( C ) Representative images of the TUNEL assay were used to confirm the induction of apoptosis in the liver sections (Scale bars: 50 μm, Magnification: 20×). ( D ) Primary hepatocytes were isolated from WT and Klf 10 KO mice. The CCK-8 assay determined the cell viability of primary hepatocytes following treatment with 5 mM glucose, 25 mM glucose, 25 mM glucose/fructose, or 25 mM glucose with 0.5 mM palmitic acid for 24 h. * p < 0.05 vs. 5 mM glucose group. # p < 0.05 vs. treatment-matched. All data are representative of at least three independent experiments and expressed as mean ± SEM. Charts were produced using GraphPad Prism 5.0. Statistical differences were determined by two-way ANOVA with Mann–Whitney U test using SPSS v17.0. WT, wild type; KO, knockout; CD, control chow diet; HSD, high-sucrose diet, CCK-8, Cell Counting Kit-8.

Article Snippet: The TransDetect ® In Situ Fluorescein TUNEL Apoptosis Detection Kit was purchased from Transgen Biotech (Beijing, China) and used according to the manufacturer’s instructions.

Techniques: Control, Western Blot, TUNEL Assay, Isolation, CCK-8 Assay, Produced, MANN-WHITNEY, Knock-Out, Cell Counting

Journal: Cells

Article Title: HOXA9 Transcriptionally Promotes Apoptosis and Represses Autophagy by Targeting NF-κB in Cutaneous Squamous Cell Carcinoma

doi: 10.3390/cells8111360

Figure Lengend Snippet: Transcriptomic analysis of HOXA9-regulated genes by RNA-Seq in cutaneous squamous cell carcinoma (cSCC) cells. Over-represented categories by GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of differently-expressed genes. BP: biological process. The “NF-kappaB signaling pathway”, “Apoptosis” and “Regulation of autophagy” were highlighted.

Article Snippet: Cells on 60-mm dishes were transfected with siNC, siHOXA9_01, or siHOXA9_02 and cultivated for 36 h. Cell apoptosis was detected using TransDetect Annexin V-FITC/PI cell apoptosis detection kit (TransGen Biotech, Beijing, China) and quantified by flow cytometry using AQUIOS CL Flow Cytometer (Beckman Coulter, Brea, CA, USA).

Techniques: Activity Assay

Journal: Cells

Article Title: HOXA9 Transcriptionally Promotes Apoptosis and Represses Autophagy by Targeting NF-κB in Cutaneous Squamous Cell Carcinoma

doi: 10.3390/cells8111360

Figure Lengend Snippet: HOXA9 represses cell proliferation while promotes apoptosis in cSCC cells. ( a ) HOXA9 protein expression was detected after knockdown of HOXA9 by siRNAs in cSCC cells. Measurements of cell proliferation by CCK-8 assay ( n = 3) ( b ) and apoptosis assay by Annexin V/PI double staining ( n = 3) ( c ) were performed in cSCC cells treated with siRNAs targeting HOXA9. ( d ) HOXA9 protein expression was detected by western blot after overexpression of HOXA9 in cSCC cells. Measurements of cell proliferation by CCK-8 assay ( n = 3) ( e ) and apoptosis assay by Annexin V/PI double staining ( n = 3) ( f ) were performed in cSCC cells overexpressing HOXA9. ( g ) The autophagy in cSCC cells following HOXA9 knockdown was evaluated by LC3 staining. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells on 60-mm dishes were transfected with siNC, siHOXA9_01, or siHOXA9_02 and cultivated for 36 h. Cell apoptosis was detected using TransDetect Annexin V-FITC/PI cell apoptosis detection kit (TransGen Biotech, Beijing, China) and quantified by flow cytometry using AQUIOS CL Flow Cytometer (Beckman Coulter, Brea, CA, USA).

Techniques: Expressing, CCK-8 Assay, Apoptosis Assay, Double Staining, Western Blot, Over Expression, Staining

Journal: Cells

Article Title: HOXA9 Transcriptionally Promotes Apoptosis and Represses Autophagy by Targeting NF-κB in Cutaneous Squamous Cell Carcinoma

doi: 10.3390/cells8111360

Figure Lengend Snippet: Loss of HOXA9 inhibited apoptosis, promoted autophagy and tumor growth in vivo. ( a ) HOXA9 depletion enhances xenografts growth. Statistical data of tumor volumes represent the average of three independent experiments ± s.d, respectively. ( b ) Dissected xenografts from three sacrificed mice were imaged at the end of experiment. Black arrows point the siNC-treated xenografts while white arrows indicate siHOXA9-treated xenografts. ( c ) The expression of HOXA9 , RELA , BCL-XL , ULK1 , ATG3 , and ATG12 was detected in the dissected xenografts by qRT-PCR. Statistical data of qRT-PCR represent the average of three independent experiments ± s.d. ( d ) The protein expression levels of HOXA9, RELA (p65), BCL-XL, ULK1, ATG3, and ATG12 was detected in xenografts after siHOXA9 treatment by western blot. ( e ) Histopathology analysis (IHC staining) of HOXA9, RELA (p65), BCL-XL, ULK1, ATG3, and ATG12 on tumor sections. Scale bar, 100 µm. ( f ) A model of the HOXA9-NF-κB regulatory axis in cSCC development. In cSCC tumors, loss of HOXA9 up-regulates NF-κB and its downstream anti-apoptotic gene BCL-XL and pro-autophagic genes of ULK1 , ATG3 , and ATG12 , which contributes to the repressed apoptosis, enhanced autophagy and promotes cSCC progression. * p < 0.05, ** p < 0.01.

Article Snippet: Cells on 60-mm dishes were transfected with siNC, siHOXA9_01, or siHOXA9_02 and cultivated for 36 h. Cell apoptosis was detected using TransDetect Annexin V-FITC/PI cell apoptosis detection kit (TransGen Biotech, Beijing, China) and quantified by flow cytometry using AQUIOS CL Flow Cytometer (Beckman Coulter, Brea, CA, USA).

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Histopathology, Immunohistochemistry