Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Caffeic acid selectively eliminates teratogenic human-induced pluripotent stem cells via apoptotic cell death.

doi: 10.1016/j.phymed.2022.154144

Figure Lengend Snippet: Fig. 3. Increase of caspase activities by CAA treatment in iPSCs. (A) Apoptosis-related proteins were detected by Western blotting in CAA-treated iPSCs and hDFs. Relative band intensities were calculated after normalization to β-actin expression. (B) iPSCs and hDFs were treated with 50 and 100 μM CAA for 24 h, and the caspase-3, −8, and −9 activities were determined. Relative caspase activity compared with CAA-untreated iPSCs and hDFs was expressed as the means ± SD (n = 3). (C) iPSCs were pre-treated with or without 10 μM of caspase-3 inhibitor (z-DEVD), caspase-8 inhibitor (z-IETD), caspase-9 inhibitor (z-LEHD), and pan-caspase inhibitor (z-VAD) for 30 min and then treated with 50 and 100 μM CAA. DMSO was treated as vehicle control. After 24 h, cells were stained with crystal violet solution and relative cell viability compared with CAA-untreated control iPSCs were expressed as the means ± SD (n = 3). **p < 0.01 vs. CAA-untreated control. ##p < 0.01 vs. Inhibitor-untreated control. Scale bar = 100 μM.

Article Snippet: The expression profile of human apoptosis-related proteins in CAA-treated or -untreated iPSCs was analyzed using a Proteome Profiler Human Apoptosis Array Kit A. Kim et al. Phytomedicine 102 (2022) 154144 (ARY009, R&D System, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Western Blot, Expressing, Activity Assay, Control, Staining

Journal: RSC Advances

Article Title: pH-responsive niosome-based nanocarriers of antineoplastic agents †

doi: 10.1039/d4ra01334d

Figure Lengend Snippet: Changes in expression levels of apoptosis-related proteins in T-24 cells following treatment with: curcumin (B); Tw60 : Sp60 : Chol – S4 (C); Tw60 : Sp60 : Chol : HD-PAA 17 – S19 (D), as compared to untreated control (A). Cells were exposed to equieffective concentrations (IC 50 ) of free curcumin and its niosome formulations for 24 h, following which a human proteome profiler assay was performed according to manufacturer's instructions. Further densitometric analysis of the array spots was conducted using ImageJ software and the most prominent changes in the proteome were ex-pressed graphically (E). Legend: 1 – bad; 2 – TRAILR1, 3 – TRAILR2; 4 – bcl-2; 5 – bcl- x ; 6 – fas, CD95; 7 – pro-caspase 3; 8 – HIF-1α; 9 – HMOX1 (antiapoptotic); 10 – cIAP1; 11 – HSP60; 12 – survivin; 13 – claspin; 14 – HSP70; 15 – TNF RI; 16 – XIAP; 17 – HTRA/Omi.

Article Snippet: Proteome Profiler Human Apoptosis Array Kit, R&D Systems and a Proteome Profiler Human Cytokine Array Kit, R&D Systems were used for the parallel determination of multiple apoptosis- and inflammation-related protein endpoints.

Techniques: Expressing, Control, Software

Journal: RSC Advances

Article Title: pH-responsive niosome-based nanocarriers of antineoplastic agents †

doi: 10.1039/d4ra01334d

Figure Lengend Snippet: Changes in expression levels of apoptosis-related proteins in MJ cells following treatment with: curcumin (B); Tw60 : Sp60 : Chol – S4 (C); Tw60 : Sp60 : Ch : HD-PAA 17 – S19 (D), as compared to untreated control (A). Cells were exposed to equieffective concentrations (IC 50 ) of free curcumin and its niosome formulations for 24 h, following which a human proteome profiler assay was performed according to manufacturer's instructions. Further densitometric analysis of the array spots was conducted using ImageJ software and the most prominent changes in the proteome were ex-pressed graphically (E). Legend: 1 – bcl-2; 2 – bcl- x ; 3 – HIF-1α; 4 – phospho p53; 5 – cIAP1; 6 – cIAP2; 7 – survivin; 8 – claspin; 9 – HSP70; 10 – XIAP.

Article Snippet: Proteome Profiler Human Apoptosis Array Kit, R&D Systems and a Proteome Profiler Human Cytokine Array Kit, R&D Systems were used for the parallel determination of multiple apoptosis- and inflammation-related protein endpoints.

Techniques: Expressing, Control, Software

Journal: RSC Advances

Article Title: pH-responsive niosome-based nanocarriers of antineoplastic agents †

doi: 10.1039/d4ra01334d

Figure Lengend Snippet: Changes in expression levels of inflammation-related proteins in T-24 cells following treatment with: curcumin (B); Tw60 : Sp60 : Chol – S4 (C); Tw60 : Sp60 : Ch : HD-PAA 17 – S19 (D), as compared to untreated control (A). Cells were exposed to equieffective concentrations (IC 50 ) of free curcumin and its niosome formulations for 24 h, following which a human cytokine profiler assay was performed according to manufacturer instructions. Further densitometric analysis of the array spots was conducted using ImageJ software and the most prominent changes in the proteome were expressed graphically (E). Legend: 1 – MIF; 2 – PAI1; 3 – G-CSF; 4 – ICAM-1; 5 – IL-6; 6 – IL-18; 7 – IL-8; 8 – CXCL1; 9 – IL-1α; 10 – IL-1β.

Article Snippet: Proteome Profiler Human Apoptosis Array Kit, R&D Systems and a Proteome Profiler Human Cytokine Array Kit, R&D Systems were used for the parallel determination of multiple apoptosis- and inflammation-related protein endpoints.

Techniques: Expressing, Control, Software

Journal: RSC Advances

Article Title: pH-responsive niosome-based nanocarriers of antineoplastic agents †

doi: 10.1039/d4ra01334d

Figure Lengend Snippet: Changes in expression levels of inflammation-related proteins in MJ cells following treatment with: curcumin (B); Tw60 : Sp60 : Ch – S4 (C); Tw60 : Sp60 : Ch : PAA 17 – S19 (D), as compared to untreated control (A). Cells were exposed to equieffective concentrations (IC 50 ) of free curcumin and its niosome formulations for 24 h, following which a human cytokine profiler assay was performed according to manufacturer instructions. Further densitometric analysis of the array spots was conducted using ImageJ software and the most prominent changes in the proteome were expressed graphically (E). Legend: 1 – CCL2/MCP-1; 2 – CXCL11/I-TAC; 3 – CXCL12/SDF-1; 4 – IL-13; 5 – IL-16; 6 – MIF; 7 – SERPIN E1/PAI-1; 8 – CCL5/RANTES; 9 – GM-CSF; 10 – IL-5; 11 – ICAM1/CD54; 12 – IL-6; 13 – IL-18; 14 – IL-21; 15 – IL-1α; 16 – IL-32α.

Article Snippet: Proteome Profiler Human Apoptosis Array Kit, R&D Systems and a Proteome Profiler Human Cytokine Array Kit, R&D Systems were used for the parallel determination of multiple apoptosis- and inflammation-related protein endpoints.

Techniques: Expressing, Control, Software

Journal: Drug Design, Development and Therapy

Article Title: Koenimbin, a natural dietary compound of Murraya koenigii (L) Spreng: inhibition of MCF7 breast cancer cells and targeting of derived MCF7 breast cancer stem cells (CD44+/CD24-/low): an in vitro study

doi: 10.2147/dddt.s72127

Figure Lengend Snippet: Figure 8 Quantitative analysis of the human apoptosis proteome profiler array in koenimbin-induced MCF7 cells. MCF7 cells were lysed and protein arrays were performed. Cells were treated with koenimbin 9 µg/mL for 24 hours and total cell protein was extracted. Equal amounts (300 µg) of protein from each control and treated sample were used for the assay. Quantitative analysis of the arrays showed differences in the apoptotic markers. Notes: The graph shows the difference between treated cells as well as untreated control cells (A). Representative images of the apoptotic protein array are shown for the control (B), treated (C), and the exact protein name of each dot in the array (D). The results are shown as the mean ± standard deviation for three independent experiments. *Indicates a significant difference from control (P0.05).

Article Snippet: The average intensity of 200 cells per well was quantified, and the ratios were then compared with TNF-α-stimulated, treated, and untreated cells.47 Human apoptosis proteome profiler array To investigate the pathways by which koenimbin induces apoptosis, we performed a determination of apoptosis-related proteins using a proteome profiler array (human apoptosis antibody array kit, Raybiotech, Norcross, GA, USA), accord- ing to the manufacturer’s instructions.

Techniques: Control, Protein Array, Standard Deviation

Journal: BMC Neuroscience

Article Title: Evidence for neuroprotective properties of human umbilical cord blood cells after neuronal hypoxia in vitro

doi: 10.1186/1471-2202-9-30

Figure Lengend Snippet: Long term effect of 48 hours of hypoxia on apoptosis (A-B) and necrosis (C-D) . Cultures at Day 3 after hypoxia were compared with normoxic cultures of the same age. Combined phase contrast/fluorescence micrographs of neuronal post-hypoxic cultures (A) and normoxic control cultures (B) show a conspicuous increase of apoptotic cells (annexin-V-staining, green fluorescence) and morphologic changes due to hypoxia (cell debris, retraction of dendrites). Propidium iodide (PI, red fluorescence) staining shows the influence of hypoxia (C) on the number of necrotic cells and cells in a late state of apoptosis compared to control cultures (D). In contrast to the level of apoptosis, there was no clear difference in the number necrotic cells following both culture conditions.

Article Snippet: BD Cytometric Bead Array for human apoptosis (Becton Dickinson, Erembodegem, Belgium) was used for the quantitative measurement of apoptotic proteins (cleaved PARP and Caspase-3).

Techniques: Fluorescence, Control, Staining

Journal: BMC Neuroscience

Article Title: Evidence for neuroprotective properties of human umbilical cord blood cells after neuronal hypoxia in vitro

doi: 10.1186/1471-2202-9-30

Figure Lengend Snippet: Effect of 48 hours of hypoxia on the concentration of active Caspase-3 and cleaved PARP . Data are taken in a time course of three days after hypoxia and are based on pooled lysates of the totality of SH-SY5Y cells taken out of 12 individual wells. Data are expressed in units of protein [active Caspase-3] and cleaved protein [PARP] per millilitres. Hypoxia induced apoptosis specific proteins (Caspase-3, cleaved PARP) in a time-related manner.

Article Snippet: BD Cytometric Bead Array for human apoptosis (Becton Dickinson, Erembodegem, Belgium) was used for the quantitative measurement of apoptotic proteins (cleaved PARP and Caspase-3).

Techniques: Concentration Assay

Journal: BMC Neuroscience

Article Title: Evidence for neuroprotective properties of human umbilical cord blood cells after neuronal hypoxia in vitro

doi: 10.1186/1471-2202-9-30

Figure Lengend Snippet: Effect of co-culturing of MNC on ratio of neuronal apoptosis and preservation of neuronal networks . Apoptosis was induced by 48 hours incubation of neuronal cells under hypoxic conditions. Afterwards 4.5 × 10 5 CFSE stained MNC were directly applied to neuronal cells (0.3 × 10 5 /well). For three days co-cultures were observed under normoxia. In co-cultures with MNC rate of apoptosis was clearly reduced compared to post-hypoxic cultures (A). Combined phase contrast and fluorescence micrograph of post-hypoxic neuronal cells and MNC (green) in direct co-culture (B).

Article Snippet: BD Cytometric Bead Array for human apoptosis (Becton Dickinson, Erembodegem, Belgium) was used for the quantitative measurement of apoptotic proteins (cleaved PARP and Caspase-3).

Techniques: Preserving, Incubation, Staining, Fluorescence, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Association of SNPs in the PAI1 Gene with Disease Recurrence and Clinical Outcome in Bladder Cancer

doi: 10.3390/ijms24054943

Figure Lengend Snippet: PAI1 rs7242 is involved in the resistance to apoptosis induced by cisplatin. ( A ) Protein and mRNA expression of PAI1 clones in stable transfectants in UROtsa and 5637 cell lines. ( B ) Apoptosis assays using RealTime-Glo™ Annexin V Apoptosis and Caspase-Glo ® 3/7 Assays. ( C ) Profiling the expression of multiple proteins associated with apoptosis induced by cisplatin using a membrane-based sandwich immunoassay. ( D ) mRNA expression levels of ATR-CLSPN-CHEK1 pathway. ( E ) Silencing CLSPN by siRNA in parental UROtsa and 5637 cell lines. ( F ) Cell survival assay after cisplatin treatment. ( G ) Apoptosis assay after cisplatin treatment. Bars represent SE. ** p < 0.01; *** p < 0.001, **** p < 0. 0001.

Article Snippet: To investigate how PAI1 might inhibit apoptosis, we used an array (Human Apoptosis Array Kit; ARY 009, Promega, Madison, WI, USA) which monitors 35 apoptosis-related proteins.

Techniques: Expressing, Clone Assay, Membrane, Clonogenic Cell Survival Assay, Apoptosis Assay