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Millipore ddh2 o
Ddh2 O, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ddh2 o - by Bioz Stars, 2022-07
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  • 97
    Millipore metformin
    Lovastatin treatment induces activation of LKB1 and AMPK in SCC cells. (A and B) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in SCC25 treated with known activators of this pathway, <t>metformin</t> and AICR, and an inhibitor compound C (CC). ( C and D) Similar Western blot analyses of treatments of SCC9 and SCC25 cells with 0–25 µM lovastatin for 24 hrs demonstrated a dose dependant induction of activated LKB1 and AMPK.
    Metformin, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
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    94
    Millipore dids
    Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures ). A : A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM <t>NS3728</t> or 400 μM <t>DIDS.</t> Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B : A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t -test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D : A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).
    Dids, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore thioflavin s solution
    Transthyretin (TTR)-positive staining associates with Thioflavin S-positivity in human calcified aortic valves. Representative ( A ) β-amyloid (Aβ), ( B ) TTR, ( C , D ) Congo Red (CR—bright field, CR pol —polarized light) and ( E ) <t>Thioflavin</t> S (Thio S) fluorescence images of human calcified aortic valves (decalcified) of patient with aortic stenosis. One donor out of n = 9 is shown. ( G ) Positive control for TTR (amyloid within the ligamentum flavum). ( F , H – J ) Brain tissue with cerebral amyloid angiopathy as positive control for Aβ ( F ), Congo Red ( H , I ) and Thio S ( J ). Arrow in J indicates Congo Red-positive signal showing areas with the typical “apple-green birefringence” in polarized light. Insert in D and I shows a higher magnification of the dashed area. Bar: 200 μm.
    Thioflavin S Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore acrylamide
    <t>Acrylamide</t> downregulated the expression of PTEN and upregulated the expression of p-AKT in HepG2 cells. A and B: The expression of PTEN, p-AKT, cyclin1, EGFR and CYP2E1 after treatment with acrylamide (0 ¦̭ol/L, 50 ¦̭ol/L, 100 ¦̭ol/L, 500 ¦̭ol/L) for 24 hours. C: The expression of cyclin D1, p-AKTafter treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination. A, B and C, as tested by Western blotting. D and E: The surviving fraction and apoptosis rate of HepG2 cells after treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination, as measured by MTT assay and flow cytometry, respectively. *P
    Acrylamide, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Lovastatin treatment induces activation of LKB1 and AMPK in SCC cells. (A and B) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in SCC25 treated with known activators of this pathway, metformin and AICR, and an inhibitor compound C (CC). ( C and D) Similar Western blot analyses of treatments of SCC9 and SCC25 cells with 0–25 µM lovastatin for 24 hrs demonstrated a dose dependant induction of activated LKB1 and AMPK.

    Journal: PLoS ONE

    Article Title: Lovastatin Induces Multiple Stress Pathways Including LKB1/AMPK Activation That Regulate Its Cytotoxic Effects in Squamous Cell Carcinoma Cells

    doi: 10.1371/journal.pone.0046055

    Figure Lengend Snippet: Lovastatin treatment induces activation of LKB1 and AMPK in SCC cells. (A and B) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in SCC25 treated with known activators of this pathway, metformin and AICR, and an inhibitor compound C (CC). ( C and D) Similar Western blot analyses of treatments of SCC9 and SCC25 cells with 0–25 µM lovastatin for 24 hrs demonstrated a dose dependant induction of activated LKB1 and AMPK.

    Article Snippet: Cells were also treated with AMPK activators, 0–20 mM metformin (Sigma, 500 mM stock in ddH2 O) and 0–1 mM AICR (Sigma, 100 mM stock in ddH2 O); and 0–5 µM AMPK inhibitor, compound C (Calbiochem, San Diego, CA, USA, 10 mM stock in DMSO).

    Techniques: Activation Assay, Western Blot

    Deletion of LKB1 in MEFs inhibits lovastatin-induced cytotoxicity. (A and B) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in wild-type LKB1+/+ murine embryonic fibroblasts (MEFs) compared to LKB1−/− MEFs. Lovastatin treatment (24 hr- 0, 1, 10 and 25 µM) induced phosphorylation of both LKB1 and AMPK only in the LKB1+/+ MEFs and were undetected in the LKB1−/− MEFs (due to the lack of expression of LKB1, actin was used as a loading control in the LKB1−/− MEFs). (C) MTT cell viability assay of LKB1+/+ and LKB1−/− MEFs treated with a range of lovastatin concentrations of up to 25 µM for 48 and 72 hrs. LKB1+/+ MEFs were significantly more sensitive to 48 hr lovastatin treatments than the LKB1−/−MEFs. However, at the 72 hr time point, the difference in cytotoxicity observed within these two sets of MEFs was not evident. (D) Fluorescent microscopic localization of exogenously expressed green fluorescent protein (GFP)/LKB1 chimeric protein in the LKB1−/− MEFs. In the control cells, LKB1 was exclusively localized in the nucleus of expressing LKB1−/− cells, while in the lovastatin treated cells greater than 80% of expressing cells LKB1 was localized in the cytoplasm. (E) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in the human SCC cell line HeLa that is deficient in LKB1 expression. Lovastatin (24 hr- 0, 1, 10 and 25 µM) and metformin (24 hrs- 0, 1, 5, 10 mM) failed to induce the phosphorylation of AMPK (due to the lack of expression of LKB1, actin was used as a loading control).

    Journal: PLoS ONE

    Article Title: Lovastatin Induces Multiple Stress Pathways Including LKB1/AMPK Activation That Regulate Its Cytotoxic Effects in Squamous Cell Carcinoma Cells

    doi: 10.1371/journal.pone.0046055

    Figure Lengend Snippet: Deletion of LKB1 in MEFs inhibits lovastatin-induced cytotoxicity. (A and B) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in wild-type LKB1+/+ murine embryonic fibroblasts (MEFs) compared to LKB1−/− MEFs. Lovastatin treatment (24 hr- 0, 1, 10 and 25 µM) induced phosphorylation of both LKB1 and AMPK only in the LKB1+/+ MEFs and were undetected in the LKB1−/− MEFs (due to the lack of expression of LKB1, actin was used as a loading control in the LKB1−/− MEFs). (C) MTT cell viability assay of LKB1+/+ and LKB1−/− MEFs treated with a range of lovastatin concentrations of up to 25 µM for 48 and 72 hrs. LKB1+/+ MEFs were significantly more sensitive to 48 hr lovastatin treatments than the LKB1−/−MEFs. However, at the 72 hr time point, the difference in cytotoxicity observed within these two sets of MEFs was not evident. (D) Fluorescent microscopic localization of exogenously expressed green fluorescent protein (GFP)/LKB1 chimeric protein in the LKB1−/− MEFs. In the control cells, LKB1 was exclusively localized in the nucleus of expressing LKB1−/− cells, while in the lovastatin treated cells greater than 80% of expressing cells LKB1 was localized in the cytoplasm. (E) Western blot analysis of activated phosphorylated LKB1 (ser428) and AMPK (thr172) along with total LKB1 and AMPK as loading controls in the human SCC cell line HeLa that is deficient in LKB1 expression. Lovastatin (24 hr- 0, 1, 10 and 25 µM) and metformin (24 hrs- 0, 1, 5, 10 mM) failed to induce the phosphorylation of AMPK (due to the lack of expression of LKB1, actin was used as a loading control).

    Article Snippet: Cells were also treated with AMPK activators, 0–20 mM metformin (Sigma, 500 mM stock in ddH2 O) and 0–1 mM AICR (Sigma, 100 mM stock in ddH2 O); and 0–5 µM AMPK inhibitor, compound C (Calbiochem, San Diego, CA, USA, 10 mM stock in DMSO).

    Techniques: Western Blot, Expressing, MTT Assay, Viability Assay

    Lovastatin but not metformin can enhance the cytotoxic effects of gefitinib in LKB1 deficient tumour cells. (A and B) MTT cell viability assays to evaluate the potential of metformin (0–20 mM) or lovastatin (0–100 µM) treatments to enhance the cytotoxicity of gefitinib (1–25 µM) in various LKB1 deficient cell lines including A549 (NSCLC) and HeLa and LKB1 expressing SCC25 and SCC9 cell lines. Metformin and lovastatin treatments were for 72 hrs (24 hr pretreatment) while the addition of gefitinib was for 48 hrs. (C and D) The Chou-Talalay method was used to distinguish between antagonistic, additive and synergistic cytotoxicity in the A549 and SCC9 with the combination of agents at various fractions of cell cytotoxicity (Fractional Effect). The Combination Index (CI) values of 1 are additive, less than 1 are synergistic and more than 1 are antagonistic and determined by CalcuSyn 2.0 software analysis. In the LKB1 deficient A549 cell line, metformin in combination with 10 µM gefitinib displayed antagonism in combination, while lovastatin in combination with 10 µM gefitinib consistently displayed synergism while in the SCC9 cell lines similar combinations of metformin or lovastatin with 10 µM gefitinib both displayed synergy in combination.

    Journal: PLoS ONE

    Article Title: Lovastatin Induces Multiple Stress Pathways Including LKB1/AMPK Activation That Regulate Its Cytotoxic Effects in Squamous Cell Carcinoma Cells

    doi: 10.1371/journal.pone.0046055

    Figure Lengend Snippet: Lovastatin but not metformin can enhance the cytotoxic effects of gefitinib in LKB1 deficient tumour cells. (A and B) MTT cell viability assays to evaluate the potential of metformin (0–20 mM) or lovastatin (0–100 µM) treatments to enhance the cytotoxicity of gefitinib (1–25 µM) in various LKB1 deficient cell lines including A549 (NSCLC) and HeLa and LKB1 expressing SCC25 and SCC9 cell lines. Metformin and lovastatin treatments were for 72 hrs (24 hr pretreatment) while the addition of gefitinib was for 48 hrs. (C and D) The Chou-Talalay method was used to distinguish between antagonistic, additive and synergistic cytotoxicity in the A549 and SCC9 with the combination of agents at various fractions of cell cytotoxicity (Fractional Effect). The Combination Index (CI) values of 1 are additive, less than 1 are synergistic and more than 1 are antagonistic and determined by CalcuSyn 2.0 software analysis. In the LKB1 deficient A549 cell line, metformin in combination with 10 µM gefitinib displayed antagonism in combination, while lovastatin in combination with 10 µM gefitinib consistently displayed synergism while in the SCC9 cell lines similar combinations of metformin or lovastatin with 10 µM gefitinib both displayed synergy in combination.

    Article Snippet: Cells were also treated with AMPK activators, 0–20 mM metformin (Sigma, 500 mM stock in ddH2 O) and 0–1 mM AICR (Sigma, 100 mM stock in ddH2 O); and 0–5 µM AMPK inhibitor, compound C (Calbiochem, San Diego, CA, USA, 10 mM stock in DMSO).

    Techniques: MTT Assay, Expressing, Software

    Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures ). A : A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B : A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t -test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D : A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Downregulation of LRRC8A protects human ovarian and alveolar carcinoma cells against Cisplatin-induced expression of p53, MDM2, p21Waf1/Cip1, and Caspase-9/-3 activation

    doi: 10.1152/ajpcell.00256.2015

    Figure Lengend Snippet: Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures ). A : A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B : A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t -test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D : A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).

    Article Snippet: The experiment was run for the total of 30 min, in the absence or presence of inhibitors, i.e., the specific VRAC/VSOAC blocker NS3728 (a gift from NeuroSearch, Denmark) and DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulfonate, dissolved in ddH2 O, Sigma Aldrich).

    Techniques: Activation Assay, Caspase-3 Activity Assay

    Impairment of swelling induced taurine release following pharmacological inhibition or transient knock-down of LRRC8A in wild-type A2780 cells. Volume-sensitive taurine release and LRRC8A protein expression were measured in ovarian A2780WT cells by tracer technique and Western blot analysis, respectively. A2780WT cells were loaded with [ 3 H]taurine for 2 h, washed, and exposed to isosmotic NaCl medium (300 mOsM) for 10 min and subsequently to hyposmotic medium (200 mOsM) for 20 min (arrow in A indicates shift to hypotonicity). Samples were taken every second minute. A : fractional rate constant (min −1 ) for taurine release was determined and plotted vs. time (min) under isosmotic and hyposmotic conditions in the absence (○) or presence of the VRAC/VSOAC blockers NS3728 (●) or DIDS (■). The anion blockers NS3728 and DIDS were used in the concentration of 10 μM (≈2.5 μM free) and 100 μM, respectively, and present during the isotonic/hypotonic release experiment. Data represent 1 of 3 sets of experiments. B : maximal rate constant for taurine release was determined 6 min after hyposmotic exposure as illustrated in A . Data represent the mean maximal rate constants determined from 3 sets of experiments ± SE. *Significantly reduced compared with control (Student's t -test). C : efficiency of siRNA-mediated LRRC8A knock-down in A2780WT cells was determined by Western blot analysis using control, scramble siRNA and LRRC8A siRNA transfected cells and specific monoclonal antibody raised against human LRRC8A and β-actin (loading control). D : LRRC8A/β-actin protein band intensity ratios were calculated from Western blots shown in C and values given relative to control cells. Data represent mean of 4 experiments ± SE. * and #: Significantly reduced compared with Control and Scramble siRNA, respectively (Student's t -test). Scramble siRNA was not significantly larger than Control ( P = 0.15, Student's t -test). E : maximal rate constant for taurine release was determined at time 6 min after hypotonic exposure in A2780WT (open bar) cells or in A2780 cells exposed to scramble siRNA (light gray bar) or siRNA directed against human LRRC8A (dark gray bar). Data represent mean of 4 experiments ± SE. *Significantly reduced compared with Control (Student's t -test).

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Downregulation of LRRC8A protects human ovarian and alveolar carcinoma cells against Cisplatin-induced expression of p53, MDM2, p21Waf1/Cip1, and Caspase-9/-3 activation

    doi: 10.1152/ajpcell.00256.2015

    Figure Lengend Snippet: Impairment of swelling induced taurine release following pharmacological inhibition or transient knock-down of LRRC8A in wild-type A2780 cells. Volume-sensitive taurine release and LRRC8A protein expression were measured in ovarian A2780WT cells by tracer technique and Western blot analysis, respectively. A2780WT cells were loaded with [ 3 H]taurine for 2 h, washed, and exposed to isosmotic NaCl medium (300 mOsM) for 10 min and subsequently to hyposmotic medium (200 mOsM) for 20 min (arrow in A indicates shift to hypotonicity). Samples were taken every second minute. A : fractional rate constant (min −1 ) for taurine release was determined and plotted vs. time (min) under isosmotic and hyposmotic conditions in the absence (○) or presence of the VRAC/VSOAC blockers NS3728 (●) or DIDS (■). The anion blockers NS3728 and DIDS were used in the concentration of 10 μM (≈2.5 μM free) and 100 μM, respectively, and present during the isotonic/hypotonic release experiment. Data represent 1 of 3 sets of experiments. B : maximal rate constant for taurine release was determined 6 min after hyposmotic exposure as illustrated in A . Data represent the mean maximal rate constants determined from 3 sets of experiments ± SE. *Significantly reduced compared with control (Student's t -test). C : efficiency of siRNA-mediated LRRC8A knock-down in A2780WT cells was determined by Western blot analysis using control, scramble siRNA and LRRC8A siRNA transfected cells and specific monoclonal antibody raised against human LRRC8A and β-actin (loading control). D : LRRC8A/β-actin protein band intensity ratios were calculated from Western blots shown in C and values given relative to control cells. Data represent mean of 4 experiments ± SE. * and #: Significantly reduced compared with Control and Scramble siRNA, respectively (Student's t -test). Scramble siRNA was not significantly larger than Control ( P = 0.15, Student's t -test). E : maximal rate constant for taurine release was determined at time 6 min after hypotonic exposure in A2780WT (open bar) cells or in A2780 cells exposed to scramble siRNA (light gray bar) or siRNA directed against human LRRC8A (dark gray bar). Data represent mean of 4 experiments ± SE. *Significantly reduced compared with Control (Student's t -test).

    Article Snippet: The experiment was run for the total of 30 min, in the absence or presence of inhibitors, i.e., the specific VRAC/VSOAC blocker NS3728 (a gift from NeuroSearch, Denmark) and DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulfonate, dissolved in ddH2 O, Sigma Aldrich).

    Techniques: Inhibition, Expressing, Western Blot, Concentration Assay, Transfection

    Administration of anion- and cation-channel blockers reduces Cisplatin-induced expression of p53 and p21 Waf1/Cip1 in wild-type A2780 cells (A2780WT) to the same level as observed in Cisplatin-resistant A2780 cells (A2780CisR). Whole cell protein lysates for Western blot analysis were obtained from A2780WT and A2780CisR cells exposed to 10 μM Cisplatin for 18 h in combination with the anion/amino acid channel blockers NS3728 (100 μM)/DIDS (400 μM) or the cation-channel (TASK-2) blocker clofilium (25 μM). Only A2780WT cells were exposed to channel blockers. Protein expression of LRRC8A, p53, p-p53 (Ser15), p21, and Bax was determined using specific antibodies (see experimental procedures ). β-Actin was used as loading control. A : representative Western blot. B : p53, pp53, p21, and Bax protein expression relative to β-actin in A2780WT and A2780CisR (open bars) control cells and following Cisplatin exposure (black bars). Data represent 4–9 individual sets of experiments where ratios are given relative to the untreated WT cells ± SE. * and #: Expression is significantly increased in Cisplatin-treated cell compared with control cells and significantly reduced in Cisplatin-treated A2780CisR cells compared with Cisplatin-treated A2780WT cells, respectively (ANOVA, Fisher LSD method). C : graph represents the relative effect of Cisplatin (Cisplatin-treated/respective untreated control) on LRRC8A, p53, p-p53, and p21 protein expression in A2780WT, A2780CisR (CisR), and A2780WT cells cotreated with the anion- and cation-channel blockers. Data represent mean of 4–9 individual experiments ± SE. *Significantly reduced effect of Cisplatin compared with A2780WT (ANOVA, Fisher LSD method).

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Downregulation of LRRC8A protects human ovarian and alveolar carcinoma cells against Cisplatin-induced expression of p53, MDM2, p21Waf1/Cip1, and Caspase-9/-3 activation

    doi: 10.1152/ajpcell.00256.2015

    Figure Lengend Snippet: Administration of anion- and cation-channel blockers reduces Cisplatin-induced expression of p53 and p21 Waf1/Cip1 in wild-type A2780 cells (A2780WT) to the same level as observed in Cisplatin-resistant A2780 cells (A2780CisR). Whole cell protein lysates for Western blot analysis were obtained from A2780WT and A2780CisR cells exposed to 10 μM Cisplatin for 18 h in combination with the anion/amino acid channel blockers NS3728 (100 μM)/DIDS (400 μM) or the cation-channel (TASK-2) blocker clofilium (25 μM). Only A2780WT cells were exposed to channel blockers. Protein expression of LRRC8A, p53, p-p53 (Ser15), p21, and Bax was determined using specific antibodies (see experimental procedures ). β-Actin was used as loading control. A : representative Western blot. B : p53, pp53, p21, and Bax protein expression relative to β-actin in A2780WT and A2780CisR (open bars) control cells and following Cisplatin exposure (black bars). Data represent 4–9 individual sets of experiments where ratios are given relative to the untreated WT cells ± SE. * and #: Expression is significantly increased in Cisplatin-treated cell compared with control cells and significantly reduced in Cisplatin-treated A2780CisR cells compared with Cisplatin-treated A2780WT cells, respectively (ANOVA, Fisher LSD method). C : graph represents the relative effect of Cisplatin (Cisplatin-treated/respective untreated control) on LRRC8A, p53, p-p53, and p21 protein expression in A2780WT, A2780CisR (CisR), and A2780WT cells cotreated with the anion- and cation-channel blockers. Data represent mean of 4–9 individual experiments ± SE. *Significantly reduced effect of Cisplatin compared with A2780WT (ANOVA, Fisher LSD method).

    Article Snippet: The experiment was run for the total of 30 min, in the absence or presence of inhibitors, i.e., the specific VRAC/VSOAC blocker NS3728 (a gift from NeuroSearch, Denmark) and DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulfonate, dissolved in ddH2 O, Sigma Aldrich).

    Techniques: Expressing, Western Blot

    Transthyretin (TTR)-positive staining associates with Thioflavin S-positivity in human calcified aortic valves. Representative ( A ) β-amyloid (Aβ), ( B ) TTR, ( C , D ) Congo Red (CR—bright field, CR pol —polarized light) and ( E ) Thioflavin S (Thio S) fluorescence images of human calcified aortic valves (decalcified) of patient with aortic stenosis. One donor out of n = 9 is shown. ( G ) Positive control for TTR (amyloid within the ligamentum flavum). ( F , H – J ) Brain tissue with cerebral amyloid angiopathy as positive control for Aβ ( F ), Congo Red ( H , I ) and Thio S ( J ). Arrow in J indicates Congo Red-positive signal showing areas with the typical “apple-green birefringence” in polarized light. Insert in D and I shows a higher magnification of the dashed area. Bar: 200 μm.

    Journal: Cells

    Article Title: Integrative Multi-Omics Analysis in Calcific Aortic Valve Disease Reveals a Link to the Formation of Amyloid-Like Deposits

    doi: 10.3390/cells9102164

    Figure Lengend Snippet: Transthyretin (TTR)-positive staining associates with Thioflavin S-positivity in human calcified aortic valves. Representative ( A ) β-amyloid (Aβ), ( B ) TTR, ( C , D ) Congo Red (CR—bright field, CR pol —polarized light) and ( E ) Thioflavin S (Thio S) fluorescence images of human calcified aortic valves (decalcified) of patient with aortic stenosis. One donor out of n = 9 is shown. ( G ) Positive control for TTR (amyloid within the ligamentum flavum). ( F , H – J ) Brain tissue with cerebral amyloid angiopathy as positive control for Aβ ( F ), Congo Red ( H , I ) and Thio S ( J ). Arrow in J indicates Congo Red-positive signal showing areas with the typical “apple-green birefringence” in polarized light. Insert in D and I shows a higher magnification of the dashed area. Bar: 200 μm.

    Article Snippet: For Thioflavin S staining, hydrated paraffin sections were stained for 5 min in Mayer′s hemalum solution, washed with water for 5 min and then stained with 1% Thioflavin S solution (w /v in ddH2 O; Sigma, Munich, Germany) for 5 min. Staining was differentiated in 70% ethanol, rinsed in ddH2O and mounted in glycerol-gelatin.

    Techniques: Staining, Fluorescence, Positive Control

    Acrylamide downregulated the expression of PTEN and upregulated the expression of p-AKT in HepG2 cells. A and B: The expression of PTEN, p-AKT, cyclin1, EGFR and CYP2E1 after treatment with acrylamide (0 ¦̭ol/L, 50 ¦̭ol/L, 100 ¦̭ol/L, 500 ¦̭ol/L) for 24 hours. C: The expression of cyclin D1, p-AKTafter treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination. A, B and C, as tested by Western blotting. D and E: The surviving fraction and apoptosis rate of HepG2 cells after treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination, as measured by MTT assay and flow cytometry, respectively. *P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: Acrylamide downregulated the expression of PTEN and upregulated the expression of p-AKT in HepG2 cells. A and B: The expression of PTEN, p-AKT, cyclin1, EGFR and CYP2E1 after treatment with acrylamide (0 ¦̭ol/L, 50 ¦̭ol/L, 100 ¦̭ol/L, 500 ¦̭ol/L) for 24 hours. C: The expression of cyclin D1, p-AKTafter treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination. A, B and C, as tested by Western blotting. D and E: The surviving fraction and apoptosis rate of HepG2 cells after treatment with acrylamide (100 ¦̭ol/L), LY294002 (20 ¦̭ol/L), and the combination, as measured by MTT assay and flow cytometry, respectively. *P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, MTT Assay, Flow Cytometry, Cytometry

    Curcumin suppressed the upregulation of miR-21 in HepG2 cells induced by acrylamide and induced cell apoptosis. A: Apoptosis rate of HepG2 cells after treatment with curcumin (10 ¦̭ol/L) for 24 hours, as measured by flow cytometry analysis. B: HepG2 cell apoptosis after treatment with curcumin (0 ¦̭ol/L, 1 ¦̭ol/L, 5 ¦̭ol/L, 10 ¦̭ol/L) for 24 hours, as assessed by Hoechest 33258 staining. C and D: The level of PTEN, p-AKT, Bcl2 and Bax in HepG2 cells after pre-treatment with curcumin (10 ¦̭ol/L) for 2 hours and combination with acrylamide for 24 hours, as analyzed by western blotting. E: The expression of miR-21 in HepG2 cells after treatment with curcumin (0 ¦̭ol/L, 1 ¦̭ol/L, 5 ¦̭ol/L, 10 ¦̭ol/L) for 24 hours, as detected by qRT-PCR. F: The expression of miR-21 in HepG2 cells, after pre-treatment by curcumin (10 ¦̭ol/L) for 2 hours and combination with acrylamide for 24 hours, as detected by qRT-PCR. * P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: Curcumin suppressed the upregulation of miR-21 in HepG2 cells induced by acrylamide and induced cell apoptosis. A: Apoptosis rate of HepG2 cells after treatment with curcumin (10 ¦̭ol/L) for 24 hours, as measured by flow cytometry analysis. B: HepG2 cell apoptosis after treatment with curcumin (0 ¦̭ol/L, 1 ¦̭ol/L, 5 ¦̭ol/L, 10 ¦̭ol/L) for 24 hours, as assessed by Hoechest 33258 staining. C and D: The level of PTEN, p-AKT, Bcl2 and Bax in HepG2 cells after pre-treatment with curcumin (10 ¦̭ol/L) for 2 hours and combination with acrylamide for 24 hours, as analyzed by western blotting. E: The expression of miR-21 in HepG2 cells after treatment with curcumin (0 ¦̭ol/L, 1 ¦̭ol/L, 5 ¦̭ol/L, 10 ¦̭ol/L) for 24 hours, as detected by qRT-PCR. F: The expression of miR-21 in HepG2 cells, after pre-treatment by curcumin (10 ¦̭ol/L) for 2 hours and combination with acrylamide for 24 hours, as detected by qRT-PCR. * P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Flow Cytometry, Cytometry, Staining, Western Blot, Expressing, Quantitative RT-PCR

    Mutual regulation of CYP2E1 and miR-21. A and B: The cell viability of HepG2 cells after the treatment with acrylamide (100 ¦̭ol/L) alone, combination with miR-21 inhibitor (100 nmol/L), siCYP2E1 alone and combination with siCYP2E1 for 24 hours, as measured by MTT assay and EdU staining. C: The expression of miR-21 after the cells were treated with acrylamide (100 ¦̭ol/L), siCYP2E1, and the combination of acrylamide and siCYP2E1 for 24 hours, as detected by qRT-PCR. D: The expression of CYP2E1 in HepG2 cells after transfection with miR-21 inhibitor (100 nmol/L) or miR-21 mimic (20 nmol/L) for 24 hours, as measured by Western blotting. * P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: Mutual regulation of CYP2E1 and miR-21. A and B: The cell viability of HepG2 cells after the treatment with acrylamide (100 ¦̭ol/L) alone, combination with miR-21 inhibitor (100 nmol/L), siCYP2E1 alone and combination with siCYP2E1 for 24 hours, as measured by MTT assay and EdU staining. C: The expression of miR-21 after the cells were treated with acrylamide (100 ¦̭ol/L), siCYP2E1, and the combination of acrylamide and siCYP2E1 for 24 hours, as detected by qRT-PCR. D: The expression of CYP2E1 in HepG2 cells after transfection with miR-21 inhibitor (100 nmol/L) or miR-21 mimic (20 nmol/L) for 24 hours, as measured by Western blotting. * P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: MTT Assay, Staining, Expressing, Quantitative RT-PCR, Transfection, Western Blot

    MiR-21 inhibitor reversed HepG2 cell proliferation induced by acrylamide. A and B: Level of miR-21 and the rate of colony formation after the cells were transfected with miR-21 inhibitor (100 nmol/L), as analyzed by qRT-PCR and colony formation assay. C: HepG2 cell proliferation after treatment with acrylamide (100 ¦̭ol/L), miR-21 inhibitor (100 nmol/L) and the combination, as analyzed by EdU staining assay. D: The level of miR- 21 after treatment with miR-21 inhibitor (100 nmol/L) and acrylamide (100 ¦̭ol/L) in HepG2 cells. * P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: MiR-21 inhibitor reversed HepG2 cell proliferation induced by acrylamide. A and B: Level of miR-21 and the rate of colony formation after the cells were transfected with miR-21 inhibitor (100 nmol/L), as analyzed by qRT-PCR and colony formation assay. C: HepG2 cell proliferation after treatment with acrylamide (100 ¦̭ol/L), miR-21 inhibitor (100 nmol/L) and the combination, as analyzed by EdU staining assay. D: The level of miR- 21 after treatment with miR-21 inhibitor (100 nmol/L) and acrylamide (100 ¦̭ol/L) in HepG2 cells. * P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Transfection, Quantitative RT-PCR, Colony Assay, Staining

    MiR-21 regulated the signaling pathway of miR-21/ PTEN/ AKT in HepG2 cells. A and B: The expression of PTEN, p-AKT, EGFR and cyclin D1 after cells were transfected with miR-21 inhibitor (100 nmol/L) and treated with acrylamide (100 ¦̭ol/L), as determined by Western blotting. * P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: MiR-21 regulated the signaling pathway of miR-21/ PTEN/ AKT in HepG2 cells. A and B: The expression of PTEN, p-AKT, EGFR and cyclin D1 after cells were transfected with miR-21 inhibitor (100 nmol/L) and treated with acrylamide (100 ¦̭ol/L), as determined by Western blotting. * P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Expressing, Transfection, Western Blot

    Acrylamide induced miR-21 expression in HepG2 cells. A: Level of miR-21 after treatment with acrylamide (0 ¦̭ol/L, 50 ¦̭ol/L, 100 ¦̭ol/L, 500 ¦̭ol/L) for 24 hours, as analyzed by qRT-PCR. B: Level of miR-21 after treatment with acrylamide at 100 ¦̭ol/L for the indicated time points (0, 3, 6, 12, 24, and 36 hours), as tested by qRT-PCR. *P

    Journal: Journal of Biomedical Research

    Article Title: Acrylamide induces HepG2 cell proliferation through upregulation of miR-21 expression

    doi: 10.7555/JBR.31.20170016

    Figure Lengend Snippet: Acrylamide induced miR-21 expression in HepG2 cells. A: Level of miR-21 after treatment with acrylamide (0 ¦̭ol/L, 50 ¦̭ol/L, 100 ¦̭ol/L, 500 ¦̭ol/L) for 24 hours, as analyzed by qRT-PCR. B: Level of miR-21 after treatment with acrylamide at 100 ¦̭ol/L for the indicated time points (0, 3, 6, 12, 24, and 36 hours), as tested by qRT-PCR. *P

    Article Snippet: Acrylamide (purity > 99.5%, dissolved in ddH2 O in a concentration of 100 mmol/L), curcumin (purity > 80%, dissolved in DMSO in a stock concentration of 50 mmol/L), LY294002, the inhibitor of PI3K/AKT were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Expressing, Quantitative RT-PCR