Journal: Molecular medicine reports

Article Title: SRPK1‑siRNA suppresses K562 cell growth and induces apoptosis via the PARP‑caspase3 pathway.

doi: 10.3892/mmr.2017.8032

Figure Lengend Snippet: Figure 5. Western blot results showing the protein levels of caspase‑3, cleaved caspase‑3, PARP, cleaved PARP, p53 and Bcl‑2/Bax. β‑actin was used as an internal loading control. Data are representative of three separate experiments. *P<0.05, compared with control groups.

Article Snippet: The primary antibodies were partly from Cell Signaling Technology, Inc. (Beverly, MA, USA) including caspase‐3 (1:1,000), cleaved caspase‐3 (1:1,000), PARP (1:1,000), cleaved PARP (1:1,000), p53 (1:1,000), Bax (1:1,000), Bcl‐2 (1:1,000) while β-actin antibodies (1:500) were purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Techniques: Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: Effect of LPS challenge on expression of PARP1, HMGB1 and circTLK1 oxidative stress and apoptosis in HCM cells. A, The viability of HCM cells after exposure to various concentrations of LPS. B, The mRNA expression of PARP1 and HMGB1 in LPS‐stimulated HCM cells was shown. C, Western blotting for PARP1 and HMGB1 protein levels in HCM cells induced by LPS. D, The expression of circTLK1 in HCM cells. E, The production of ROS in HCM cells. The lactate dehydrogenase (LDH) (F), creatine kinase (CK) (G) levels in the cellular supernatant, and the MDA level (H) and SOD activity (I) in HCM cells were evaluated. J, The apoptosis of HCM cells was assessed. All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs control group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Expressing, Western Blot, Activity Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: CircTLK1 or PARP1 inhibition restrains LPS‐induced apoptosis in HCM cells. The mRNA (A) and protein (B) levels of PARP1 and HMGB1 in HCM cells subjected to various treatments. C, Effect of PARP1 on the acetylation of HMGB1 was evaluated by Co‐IP assay. D, The viability of HCM cells from different groups was shown. The apoptosis of HCM cells was assessed by flow cytometer (E) and TUNEL (F). G, Protein levels of Bcl‐2, Bax and cleaved caspase‐3 in HCM cells. All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Inhibition, Co-Immunoprecipitation Assay, Flow Cytometry, TUNEL Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: Effect of circTLK1 silencing or PARP1 suppression on LPS‐induced oxidative stress and mitochondrial dysfunction. A, ROS production in HCM cells. The MDA level (B) and SOD activity (C) in HCM cells were determined. D, Mitochondrial membrane potential (∆Ψm) of HCM cells was detected. E, The NAD+/NADH ratio was detected. The distribution of cytochrome C in HCM cells was determined by Western blotting (F) and immunofluorescence staining (G). All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Activity Assay, Membrane, Western Blot, Immunofluorescence, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: Effect of PARP1 or circTLK1 suppression on mtDNA damage after exposure to LPS. A, The copy number of mtDNA. B, To assess mtDNA transcript levels, mtDNA‐encoded ND1, CytB and ATP6 expression was detected. C, The 8‐OHdG level in HCM cells was determined. D, The ATP level in HCM cells was assessed. All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: Effect of circTLK1/PARP1 axis inhibition on oxidative mtDNA damage‐mediated apoptosis in LPS‐induced HCM cells. A, The apoptotic rate of HCM cells was detected. The MDA level (B) and SOD activity (C) in HCM cells were evaluated. D, The copy number of mtDNA was detected. E, The mtDNA transcript levels (ND1, CytB and ATP6) were assessed. All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Inhibition, Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: Effect of circTLK1/PARP1 axis on oxidative mtDNA damage–mediated cardiomyocyte apoptosis in septic rats. (A) and (B) PARP1 and HMGB1 expression levels in the myocardial tissues. C, The apoptosis in myocardial tissues. D, The protein levels of Bcl‐2, Bax and cleaved caspase‐3 in myocardial tissues were assessed. The levels of MDA (E) and 8‐OHdG (F) in myocardial tissues were detected. G, The mtDNA copy number in myocardial tissues was determined. H, The mtDNA transcript levels (ND1, CytB and ATP6) in myocardial tissues were determined. All results from three independent experiments were expressed as mean ± SD. * P < .05; ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: CircTLK1 promotes PARP1 expression via sponging miR‐17‐5p. A, The predicted complementarity between circTLK1 and miR‐17‐5p. B, Dual‐luciferase assay for verifying the relationship between circTLK1 and miR‐17‐5p. C, The direct binding between circTLK1 and miR‐17‐5p was confirmed by RIP assay. D, Expression of miR‐17‐5p in HCM cells was assessed. E, The mRNA expression of PARP1 in HCM cells with various treatments was detected. F, The putative region of complementarity between miR‐17‐5p and PARP1. G, The interaction between miR‐17‐5p and PARP1 was determined by dual‐luciferase assay. All results from three independent experiments were expressed as mean ± SD. ** P < .01; *** P < .001 vs the indicated group

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: Expressing, Luciferase, Binding Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: CircTLK1 modulates sepsis‐induced cardiomyocyte apoptosis via enhancing PARP1/HMGB1 axis–mediated mitochondrial DNA damage by sponging miR‐17‐5p

doi: 10.1111/jcmm.16738

Figure Lengend Snippet: A schematic overview of the role of circTLK1/miR‐17‐5p/PARP1/HMGB1 axis in LPS‐induced mtDNA oxidative damage and subsequent cardiomyocyte apoptosis

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour and incubated with primary antibodies against PARP1 (1:1000, PB9309, Boster Biological Technology), HMGB1 (1:1000, A00066‐1, Boster Biological Technology), Bcl‐2 (1:1000, A00040‐2, Boster Biological Technology), Bax (1:500, bs‐0127R, Bioss, Beijing, China), cleaved caspase‐3 (c‐Caspase3; 1:500, BA3257, Boster Biological Technology), acetylated lysine (1:500, MA1‐2021, Thermo Fisher), cytochrome C (1:500, bs‐0013R, Bioss), COX IV (1:2000, bsm‐52750R, Bioss) and GAPDH (1:1000, BA2913, Boster Biological Technology) overnight at 4°C.

Techniques: