monodansylcadaverine mdc  (Millipore)


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    Name:
    Dansylcadaverine
    Description:
    Dansylcadaverine also called as monodansylcadaverine MDC is a lysosomotropic agent It is present in acidic and lipid rich regions Dansylcadaverine also acts as a solvent polarity probe It inhibits vesicular transport and decreases the rotavirus sequestering in L cells when pre treated
    Catalog Number:
    d4008
    Price:
    None
    Applications:
    Dansylcadaverine in methanol has a reported lambdaex of 335 nm and lambdaem of 512 nm. It has been used:. as an endocytosis inhibitor in human proximal tubular HK-2 cells,(62) mouse J774A.1 macrophages and human A549 epithelial cells(63) . to label autophagic vacuoles in cardiomyocytes for fluorescence microscopy studies(64). in monodansylcadaverine (MDC) staining of fibroblasts in Ankylosing spondylitis (AS) samples(65)
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    Structured Review

    Millipore monodansylcadaverine mdc
    Dansylcadaverine
    Dansylcadaverine also called as monodansylcadaverine MDC is a lysosomotropic agent It is present in acidic and lipid rich regions Dansylcadaverine also acts as a solvent polarity probe It inhibits vesicular transport and decreases the rotavirus sequestering in L cells when pre treated
    https://www.bioz.com/result/monodansylcadaverine mdc/product/Millipore
    Average 99 stars, based on 45 article reviews
    Price from $9.99 to $1999.99
    monodansylcadaverine mdc - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "A large-scale RNA interference screen identifies genes that regulate autophagy at different stages"

    Article Title: A large-scale RNA interference screen identifies genes that regulate autophagy at different stages

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21106-5

    Monodansylcadaverine (MDC) stains autophagic K562 cells with sensitivity equivalent to LC3B immunoblotting and GFP-LC3B fluorescence microscopy. ( A ) Fluorescence microscopy. K562 cells were treated with dimethyl sulfoxide (DMSO) or 1 μM imatinib (IM) overnight followed by MDC staining. Images of live cells were taken at different exposure times (milliseconds, ms) using an inverted fluorescence microscope. Scale bar: 25 μm. ( B ) MDC Fluorescence spectrophotometry. K562 cells treated with DMSO or IM were stained with MDC. MDC fluorescence of live K562 cells was quantified using a micro-plate reader at an excitation wavelength of 335 nm and an emission wavelength of 525 nm. The relative MDC levels were obtained by dividing MDC fluorescence of IM-treated cells to that of DMSO-treated cells. Error bars represent three independent experiments. ( C ) LC3B immunoblotting. Cropped images are shown and full images are included in supplemental materials. ACTB (β actin) is the loading control. The intensities of LC3B-II or ACTB were quantified using Image J. The fold changes of LC3B-II/ACTB were obtained by dividing the ratio of LC3B-II/ACTB in IM-treated cells with that of DMSO-treated cells. ( D ) MDC staining of K562 cells treated with IM and/or bafilomycin A1 (BFA1). K562 cells were treated with a combination of 1 μM IM (10 hours) and 5 nM of BFA1 (48 hours). Cells were imaged using a fluorescence confocal microscope. ( E ) Quantification of MDC intensities. Fluorescence intensities of MDC in 10 different cells from three images were quantified using Image J. ( F ) GFP-LC3B fluorescence microscopy. K562 cells were stably transfected with a construct encoding GFP-LC3B. Cells were treated with a combination of IM and BFA1 and then imaged. ( G ) Quantification of cells with GFP-LC3B puncta. Four to five images of more than thirty K562 cells from each treatment were randomly selected for the following quantification analyses. GFP-expressing K562 cells or K562 cells with GFP puncta (indicating autophagy) were counted by three persons. Percentages of K562 cells with puncta were obtained by dividing numbers of K562 cells with GFP puncta with those of GFP-expressing cells. Scale bar: 25 nm.
    Figure Legend Snippet: Monodansylcadaverine (MDC) stains autophagic K562 cells with sensitivity equivalent to LC3B immunoblotting and GFP-LC3B fluorescence microscopy. ( A ) Fluorescence microscopy. K562 cells were treated with dimethyl sulfoxide (DMSO) or 1 μM imatinib (IM) overnight followed by MDC staining. Images of live cells were taken at different exposure times (milliseconds, ms) using an inverted fluorescence microscope. Scale bar: 25 μm. ( B ) MDC Fluorescence spectrophotometry. K562 cells treated with DMSO or IM were stained with MDC. MDC fluorescence of live K562 cells was quantified using a micro-plate reader at an excitation wavelength of 335 nm and an emission wavelength of 525 nm. The relative MDC levels were obtained by dividing MDC fluorescence of IM-treated cells to that of DMSO-treated cells. Error bars represent three independent experiments. ( C ) LC3B immunoblotting. Cropped images are shown and full images are included in supplemental materials. ACTB (β actin) is the loading control. The intensities of LC3B-II or ACTB were quantified using Image J. The fold changes of LC3B-II/ACTB were obtained by dividing the ratio of LC3B-II/ACTB in IM-treated cells with that of DMSO-treated cells. ( D ) MDC staining of K562 cells treated with IM and/or bafilomycin A1 (BFA1). K562 cells were treated with a combination of 1 μM IM (10 hours) and 5 nM of BFA1 (48 hours). Cells were imaged using a fluorescence confocal microscope. ( E ) Quantification of MDC intensities. Fluorescence intensities of MDC in 10 different cells from three images were quantified using Image J. ( F ) GFP-LC3B fluorescence microscopy. K562 cells were stably transfected with a construct encoding GFP-LC3B. Cells were treated with a combination of IM and BFA1 and then imaged. ( G ) Quantification of cells with GFP-LC3B puncta. Four to five images of more than thirty K562 cells from each treatment were randomly selected for the following quantification analyses. GFP-expressing K562 cells or K562 cells with GFP puncta (indicating autophagy) were counted by three persons. Percentages of K562 cells with puncta were obtained by dividing numbers of K562 cells with GFP puncta with those of GFP-expressing cells. Scale bar: 25 nm.

    Techniques Used: Fluorescence, Microscopy, Staining, Mass Spectrometry, Spectrophotometry, Stable Transfection, Transfection, Construct, Expressing

    2) Product Images from "Autophagy Activation Is Involved in 3,4-Methylenedioxymethamphetamine (‘Ecstasy’)—Induced Neurotoxicity in Cultured Cortical Neurons"

    Article Title: Autophagy Activation Is Involved in 3,4-Methylenedioxymethamphetamine (‘Ecstasy’)—Induced Neurotoxicity in Cultured Cortical Neurons

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0116565

    3-MA decreased MDMA-elicited autophagy activation and neurite degeneration. Cultured cortical neurons were exposed to 1 mM MDMA for 48 h in the presence or absence of 1 mM 3-MA for 48 h. Autophagy activity was assay by MDC incorporation, western blot analysis and double immunofluorescence staining with anti-LC3B and MAP2 antibodies. (A) The autophagic vacuoles were visualized by monodansylcadaverine (MDC) staining and the mean number of MDC-labeled vacuoles per cortical neurons (arrows) was quantitation. Quantitative data are expressed as mean ± S.D. n = 3. ## P
    Figure Legend Snippet: 3-MA decreased MDMA-elicited autophagy activation and neurite degeneration. Cultured cortical neurons were exposed to 1 mM MDMA for 48 h in the presence or absence of 1 mM 3-MA for 48 h. Autophagy activity was assay by MDC incorporation, western blot analysis and double immunofluorescence staining with anti-LC3B and MAP2 antibodies. (A) The autophagic vacuoles were visualized by monodansylcadaverine (MDC) staining and the mean number of MDC-labeled vacuoles per cortical neurons (arrows) was quantitation. Quantitative data are expressed as mean ± S.D. n = 3. ## P

    Techniques Used: Activation Assay, Cell Culture, Activity Assay, Western Blot, Double Immunofluorescence Staining, Staining, Labeling, Quantitation Assay

    3) Product Images from "Internalization of the opioid growth factor, [Met5]-enkephalin, is dependent on clathrin-mediated endocytosis for downregulation of cell proliferation"

    Article Title: Internalization of the opioid growth factor, [Met5]-enkephalin, is dependent on clathrin-mediated endocytosis for downregulation of cell proliferation

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    doi: 10.1152/ajpregu.00318.2010

    RhoOGF uptake is impaired by inhibitors targeting different endocytic pathways. Cells were preincubated with serum-free medium for 30 min at 37°C, and treated with monodansylcadaverine (MDC) for 1 h, EIPA for 30 min, methyl-β-cyclodextrin (MβCD) for 30 min, or both MDC and EIPA together for 30 min; some cells remained untreated [control (Ctrl)]. Both RhoOGF (10 −5 M) and transferrin (Tf; 10 μg/ml) were added to cell cultures for 30 min in the presence of inhibitors. Cells were stained with Hoechst and analyzed by Epi-F (RhoOGF, Hoechst, or Tf) or DIC microscopy. Bar = 10 μm.
    Figure Legend Snippet: RhoOGF uptake is impaired by inhibitors targeting different endocytic pathways. Cells were preincubated with serum-free medium for 30 min at 37°C, and treated with monodansylcadaverine (MDC) for 1 h, EIPA for 30 min, methyl-β-cyclodextrin (MβCD) for 30 min, or both MDC and EIPA together for 30 min; some cells remained untreated [control (Ctrl)]. Both RhoOGF (10 −5 M) and transferrin (Tf; 10 μg/ml) were added to cell cultures for 30 min in the presence of inhibitors. Cells were stained with Hoechst and analyzed by Epi-F (RhoOGF, Hoechst, or Tf) or DIC microscopy. Bar = 10 μm.

    Techniques Used: Staining, Microscopy

    4) Product Images from "Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines"

    Article Title: Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S152718

    Fluorescent microscopy demonstrating acidic vesicles via MDC staining of A549 ( Ai – Gi ) and MDA-MB-231 ( Aii – Gii ) cells treated with the compounds. Notes: DMSO-exposed cells ( Ai , Aii ) served as negative controls and displayed nonspecific MDC staining. Tamoxifen treatment ( Bi , Bii ) was used as a positive control for acidic vacuoles and displayed increased definition and intensity in the MDC-stained vacuoles. Cells treated with 2-ME ( Ci , Cii ), STX 2895 ( Di , Dii ), STX 3329 ( Ei , Eii ), STX 3450 ( Fi , Fii ) and STX 3451 ( Gi , Gii ) showed increased MDC staining indicating the potential role of autophagy in the cellular response and a decreased cell density (40× magnification, scale bar: 20 µM). Abbreviations: 2-ME, 2-methoxyestradiol; DMSO, dimethyl sulfoxide; MDC, monodansylcadaverine.
    Figure Legend Snippet: Fluorescent microscopy demonstrating acidic vesicles via MDC staining of A549 ( Ai – Gi ) and MDA-MB-231 ( Aii – Gii ) cells treated with the compounds. Notes: DMSO-exposed cells ( Ai , Aii ) served as negative controls and displayed nonspecific MDC staining. Tamoxifen treatment ( Bi , Bii ) was used as a positive control for acidic vacuoles and displayed increased definition and intensity in the MDC-stained vacuoles. Cells treated with 2-ME ( Ci , Cii ), STX 2895 ( Di , Dii ), STX 3329 ( Ei , Eii ), STX 3450 ( Fi , Fii ) and STX 3451 ( Gi , Gii ) showed increased MDC staining indicating the potential role of autophagy in the cellular response and a decreased cell density (40× magnification, scale bar: 20 µM). Abbreviations: 2-ME, 2-methoxyestradiol; DMSO, dimethyl sulfoxide; MDC, monodansylcadaverine.

    Techniques Used: Microscopy, Staining, Multiple Displacement Amplification, Positive Control

    5) Product Images from "RhoA/ROCK1 regulates Avian Reovirus S1133-induced switch from autophagy to apoptosis"

    Article Title: RhoA/ROCK1 regulates Avian Reovirus S1133-induced switch from autophagy to apoptosis

    Journal: BMC Veterinary Research

    doi: 10.1186/s12917-015-0417-6

    ARV S1133 induces autophagy and subsequent apoptosis in cultured cells. (A) DF1 cells infected with ARV S1133 at an MOI of 20. (B) Vero cells infected with ARV S1133 at an MOI of 5 for 0–42 hr. At the indicated time points, cells were stained with monodansylcadaverine (MDC) or Hoechst 332588. The percentage of positive cells was calculated for 20 independent fields at a magnification of 200×. (C) Vero cells infected with ARV S1133 at an MOI of 5 for 0–36 hr. In Hoechst 33258-stained cells (bright blue), arrows indicate apoptotic nuclei with condensed chromatin. In MDC stained cells, arrows indicate the autophagic vacuoles (×400 magnification, scale bar 10 μm).
    Figure Legend Snippet: ARV S1133 induces autophagy and subsequent apoptosis in cultured cells. (A) DF1 cells infected with ARV S1133 at an MOI of 20. (B) Vero cells infected with ARV S1133 at an MOI of 5 for 0–42 hr. At the indicated time points, cells were stained with monodansylcadaverine (MDC) or Hoechst 332588. The percentage of positive cells was calculated for 20 independent fields at a magnification of 200×. (C) Vero cells infected with ARV S1133 at an MOI of 5 for 0–36 hr. In Hoechst 33258-stained cells (bright blue), arrows indicate apoptotic nuclei with condensed chromatin. In MDC stained cells, arrows indicate the autophagic vacuoles (×400 magnification, scale bar 10 μm).

    Techniques Used: Cell Culture, Infection, Staining

    6) Product Images from "Propofol protects human keratinocytes from oxidative stress via autophagy expression"

    Article Title: Propofol protects human keratinocytes from oxidative stress via autophagy expression

    Journal: Journal of Dental Anesthesia and Pain Medicine

    doi: 10.17245/jdapm.2017.17.1.21

    Monodansylcadaverine (MDC) staining of cytoplasmic vacuoles after propofol treatment in human keratinocytes. Significant accumulation of autophagy-specific staining of MDC was observed around the nuclei in PPC/H 2 O 2 group cells.
    Figure Legend Snippet: Monodansylcadaverine (MDC) staining of cytoplasmic vacuoles after propofol treatment in human keratinocytes. Significant accumulation of autophagy-specific staining of MDC was observed around the nuclei in PPC/H 2 O 2 group cells.

    Techniques Used: Staining

    7) Product Images from "Gambogenic Acid Kills Lung Cancer Cells through Aberrant Autophagy"

    Article Title: Gambogenic Acid Kills Lung Cancer Cells through Aberrant Autophagy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0083604

    GNA increase autophagic markers in A549 and HeLa cells. A, Representative images of MDC staining. A549 cells were treated with 3 µM GNA for 24 hours, then stained with 10 µM monodansylcadaverine (MDC) and observed under a phase contrast microscope. B and C, Acridine orange staining. Cells were exposed to the indicated concentrations of GNA for 24 hours, then stained with 1 µg/ml of acridine orange (B, C) and observed under a phase contrast microscope (B) or analyzed by flow cytometry (C). Cells that were treated with 50 µg/ml of rapamycin were used as a positive control. The results shown are representative of at least 2 independent experiments. D, Representative electron microscopy images of GNA-treated A549 cells. The cells were treated with 3 µM GNA for the indicated periods of time and analyzed by electron microscopy. Arrows indicate the vacuoles contacting with vesicles that were densely filled with multi-lamellar structures.
    Figure Legend Snippet: GNA increase autophagic markers in A549 and HeLa cells. A, Representative images of MDC staining. A549 cells were treated with 3 µM GNA for 24 hours, then stained with 10 µM monodansylcadaverine (MDC) and observed under a phase contrast microscope. B and C, Acridine orange staining. Cells were exposed to the indicated concentrations of GNA for 24 hours, then stained with 1 µg/ml of acridine orange (B, C) and observed under a phase contrast microscope (B) or analyzed by flow cytometry (C). Cells that were treated with 50 µg/ml of rapamycin were used as a positive control. The results shown are representative of at least 2 independent experiments. D, Representative electron microscopy images of GNA-treated A549 cells. The cells were treated with 3 µM GNA for the indicated periods of time and analyzed by electron microscopy. Arrows indicate the vacuoles contacting with vesicles that were densely filled with multi-lamellar structures.

    Techniques Used: Staining, Microscopy, Flow Cytometry, Cytometry, Positive Control, Electron Microscopy

    8) Product Images from "Sodium formate induces autophagy and apoptosis via the JNK signaling pathway of photoreceptor cells"

    Article Title: Sodium formate induces autophagy and apoptosis via the JNK signaling pathway of photoreceptor cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2015.4675

    Sodium formate exposure induces the formation of MDC-positive autophagosomes in 661W cells. The 661W cells were exposed to (A) 0 (control), (B) 15 or (C) 30 mM sodium formate. Autophagic vacuoles were labeled with 0.05 mM MDC in phosphate-buffered saline at 37°C for 10 min. The fluorescence density and the MDC-labeled particles in the 661W cells were greater in the group subjected to sodium formate treatment for 6 h compared with the control group. MDC-labeled vesicles are indicated by arrows (Scale bar=50 µ m). MDC, monodansylcadaverine.
    Figure Legend Snippet: Sodium formate exposure induces the formation of MDC-positive autophagosomes in 661W cells. The 661W cells were exposed to (A) 0 (control), (B) 15 or (C) 30 mM sodium formate. Autophagic vacuoles were labeled with 0.05 mM MDC in phosphate-buffered saline at 37°C for 10 min. The fluorescence density and the MDC-labeled particles in the 661W cells were greater in the group subjected to sodium formate treatment for 6 h compared with the control group. MDC-labeled vesicles are indicated by arrows (Scale bar=50 µ m). MDC, monodansylcadaverine.

    Techniques Used: Labeling, Fluorescence

    9) Product Images from "A large-scale RNA interference screen identifies genes that regulate autophagy at different stages"

    Article Title: A large-scale RNA interference screen identifies genes that regulate autophagy at different stages

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21106-5

    Monodansylcadaverine (MDC) stains autophagic K562 cells with sensitivity equivalent to LC3B immunoblotting and GFP-LC3B fluorescence microscopy. ( A ) Fluorescence microscopy. K562 cells were treated with dimethyl sulfoxide (DMSO) or 1 μM imatinib (IM) overnight followed by MDC staining. Images of live cells were taken at different exposure times (milliseconds, ms) using an inverted fluorescence microscope. Scale bar: 25 μm. ( B ) MDC Fluorescence spectrophotometry. K562 cells treated with DMSO or IM were stained with MDC. MDC fluorescence of live K562 cells was quantified using a micro-plate reader at an excitation wavelength of 335 nm and an emission wavelength of 525 nm. The relative MDC levels were obtained by dividing MDC fluorescence of IM-treated cells to that of DMSO-treated cells. Error bars represent three independent experiments. ( C ) LC3B immunoblotting. Cropped images are shown and full images are included in supplemental materials. ACTB (β actin) is the loading control. The intensities of LC3B-II or ACTB were quantified using Image J. The fold changes of LC3B-II/ACTB were obtained by dividing the ratio of LC3B-II/ACTB in IM-treated cells with that of DMSO-treated cells. ( D ) MDC staining of K562 cells treated with IM and/or bafilomycin A1 (BFA1). K562 cells were treated with a combination of 1 μM IM (10 hours) and 5 nM of BFA1 (48 hours). Cells were imaged using a fluorescence confocal microscope. ( E ) Quantification of MDC intensities. Fluorescence intensities of MDC in 10 different cells from three images were quantified using Image J. ( F ) GFP-LC3B fluorescence microscopy. K562 cells were stably transfected with a construct encoding GFP-LC3B. Cells were treated with a combination of IM and BFA1 and then imaged. ( G ) Quantification of cells with GFP-LC3B puncta. Four to five images of more than thirty K562 cells from each treatment were randomly selected for the following quantification analyses. GFP-expressing K562 cells or K562 cells with GFP puncta (indicating autophagy) were counted by three persons. Percentages of K562 cells with puncta were obtained by dividing numbers of K562 cells with GFP puncta with those of GFP-expressing cells. Scale bar: 25 nm.
    Figure Legend Snippet: Monodansylcadaverine (MDC) stains autophagic K562 cells with sensitivity equivalent to LC3B immunoblotting and GFP-LC3B fluorescence microscopy. ( A ) Fluorescence microscopy. K562 cells were treated with dimethyl sulfoxide (DMSO) or 1 μM imatinib (IM) overnight followed by MDC staining. Images of live cells were taken at different exposure times (milliseconds, ms) using an inverted fluorescence microscope. Scale bar: 25 μm. ( B ) MDC Fluorescence spectrophotometry. K562 cells treated with DMSO or IM were stained with MDC. MDC fluorescence of live K562 cells was quantified using a micro-plate reader at an excitation wavelength of 335 nm and an emission wavelength of 525 nm. The relative MDC levels were obtained by dividing MDC fluorescence of IM-treated cells to that of DMSO-treated cells. Error bars represent three independent experiments. ( C ) LC3B immunoblotting. Cropped images are shown and full images are included in supplemental materials. ACTB (β actin) is the loading control. The intensities of LC3B-II or ACTB were quantified using Image J. The fold changes of LC3B-II/ACTB were obtained by dividing the ratio of LC3B-II/ACTB in IM-treated cells with that of DMSO-treated cells. ( D ) MDC staining of K562 cells treated with IM and/or bafilomycin A1 (BFA1). K562 cells were treated with a combination of 1 μM IM (10 hours) and 5 nM of BFA1 (48 hours). Cells were imaged using a fluorescence confocal microscope. ( E ) Quantification of MDC intensities. Fluorescence intensities of MDC in 10 different cells from three images were quantified using Image J. ( F ) GFP-LC3B fluorescence microscopy. K562 cells were stably transfected with a construct encoding GFP-LC3B. Cells were treated with a combination of IM and BFA1 and then imaged. ( G ) Quantification of cells with GFP-LC3B puncta. Four to five images of more than thirty K562 cells from each treatment were randomly selected for the following quantification analyses. GFP-expressing K562 cells or K562 cells with GFP puncta (indicating autophagy) were counted by three persons. Percentages of K562 cells with puncta were obtained by dividing numbers of K562 cells with GFP puncta with those of GFP-expressing cells. Scale bar: 25 nm.

    Techniques Used: Fluorescence, Microscopy, Staining, Mass Spectrometry, Spectrophotometry, Stable Transfection, Transfection, Construct, Expressing

    10) Product Images from "Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines"

    Article Title: Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S152718

    Fluorescent microscopy demonstrating acidic vesicles via MDC staining of A549 ( Ai – Gi ) and MDA-MB-231 ( Aii – Gii ) cells treated with the compounds. Notes: DMSO-exposed cells ( Ai , Aii ) served as negative controls and displayed nonspecific MDC staining. Tamoxifen treatment ( Bi , Bii ) was used as a positive control for acidic vacuoles and displayed increased definition and intensity in the MDC-stained vacuoles. Cells treated with 2-ME ( Ci , Cii ), STX 2895 ( Di , Dii ), STX 3329 ( Ei , Eii ), STX 3450 ( Fi , Fii ) and STX 3451 ( Gi , Gii ) showed increased MDC staining indicating the potential role of autophagy in the cellular response and a decreased cell density (40× magnification, scale bar: 20 µM). Abbreviations: 2-ME, 2-methoxyestradiol; DMSO, dimethyl sulfoxide; MDC, monodansylcadaverine.
    Figure Legend Snippet: Fluorescent microscopy demonstrating acidic vesicles via MDC staining of A549 ( Ai – Gi ) and MDA-MB-231 ( Aii – Gii ) cells treated with the compounds. Notes: DMSO-exposed cells ( Ai , Aii ) served as negative controls and displayed nonspecific MDC staining. Tamoxifen treatment ( Bi , Bii ) was used as a positive control for acidic vacuoles and displayed increased definition and intensity in the MDC-stained vacuoles. Cells treated with 2-ME ( Ci , Cii ), STX 2895 ( Di , Dii ), STX 3329 ( Ei , Eii ), STX 3450 ( Fi , Fii ) and STX 3451 ( Gi , Gii ) showed increased MDC staining indicating the potential role of autophagy in the cellular response and a decreased cell density (40× magnification, scale bar: 20 µM). Abbreviations: 2-ME, 2-methoxyestradiol; DMSO, dimethyl sulfoxide; MDC, monodansylcadaverine.

    Techniques Used: Microscopy, Staining, Multiple Displacement Amplification, Positive Control

    11) Product Images from "Cranberry proanthocyanidins inhibit esophageal adenocarcinoma in vitro and in vivo through pleiotropic cell death induction and PI3K/AKT/mTOR inactivation"

    Article Title: Cranberry proanthocyanidins inhibit esophageal adenocarcinoma in vitro and in vivo through pleiotropic cell death induction and PI3K/AKT/mTOR inactivation

    Journal: Oncotarget

    doi:

    Effect of C-PAC on autophagy induction in EAC cells A. Ultrastructural changes associated with C-PAC treatment of EAC Cells. Transmission electron micrographs were captured of JHAD1 and OE19 cells following 6 and 24 hours of treatment with C-PAC [50μg/ml] to evaluate autophagic vacuole formation. a) and e) show vehicle treated JHAD1 cells; whereas, i) and m) show vehicle treated OE19 cells at 6 and 24 hours, respectively. Vehicle treated cells are characterized by microvilli protruding from the cell surface, a round smoothly outlined normal appearing nuclei, normally distributed heterochromatin, well defined plasma membrane and well preserved cytoplasmic organelles. b), c), and d) show cytoplasmic vacuolization (black arrows), increased formation of single and double walled autophagic vesicles as shown as autophagosomes (blue arrowheads) and autolysomes (white arrowheads, black outline) as early as 6 hours post C-PAC treatment of JHAD1 cells. Increased autophagy continues 24 hours following C-PAC treatment as displayed in f), g), and h) with increased electron dense cargo supporting formation of degradative autophagic vacuoles. C-PAC treatment of OE19 cells induced cellular necrosis as illustrated in j) and k) by moderate chromatin clumping and nuclear disintegration or karyolysis (asterisk), accompanied by loss of plasma membrane integrity and marked loss of cellular contents as in k), l), and o) (extra-cytoplasmic cellular debris, red arrowhead). Cellular blebbing (k, n) and fragmentation of the nucleus (Karyorrhexis, black arrowhead) are evident. p) Autophagic vacuoles containing cellular debris indicate a low level of autophagy induction in OE19 cells. B. Representative photomicrographs (200X) of JHAD1 and OE33 EAC cells treated with C-PAC [50 μg/ml] for 6 and 24 hours, black arrows indicate apoptotic cells. The enlarged inset illustrates morphological changes of cytoplasmic swelling and vacuolization consistent with autophagy induction (as also noted in Figure 1D ). Monodansylcadaverine (MDC) staining and confocal microscopy were employed next to probe autophagic vacuoles in OE19, JHAD1 and OE33 cells following C-PAC treatment [50 μg/ml] for 24 hours. C-PAC treatment of OE33 cells resulted in increased accumulation of punctate staining as illustrated in large MDC positive vesicles (far right, bottom panel). C. To further investigate C-PAC induced autophagy, JHAD1, OE33 and OE19 cells were treated with C-PAC [50 μg/ml] or vehicle in triplicate and lysates collected at baseline and following 6, 24 and 48 hours of treatment; then, probed for LC3B, microtubule-associated protein 1 light chain 3 beta, an important early marker and effector of autophagy. Representative images are shown, expression values were normalized to the loading control GAPDH and a mean fold change from baseline or time of first detection calculated utilizing Quantity One software (Bio-Rad, Hercules, CA). Positive fold change values indicate increased expression and negative values reflect decreased expression. Figure 3C shows that C-PAC strongly ( P
    Figure Legend Snippet: Effect of C-PAC on autophagy induction in EAC cells A. Ultrastructural changes associated with C-PAC treatment of EAC Cells. Transmission electron micrographs were captured of JHAD1 and OE19 cells following 6 and 24 hours of treatment with C-PAC [50μg/ml] to evaluate autophagic vacuole formation. a) and e) show vehicle treated JHAD1 cells; whereas, i) and m) show vehicle treated OE19 cells at 6 and 24 hours, respectively. Vehicle treated cells are characterized by microvilli protruding from the cell surface, a round smoothly outlined normal appearing nuclei, normally distributed heterochromatin, well defined plasma membrane and well preserved cytoplasmic organelles. b), c), and d) show cytoplasmic vacuolization (black arrows), increased formation of single and double walled autophagic vesicles as shown as autophagosomes (blue arrowheads) and autolysomes (white arrowheads, black outline) as early as 6 hours post C-PAC treatment of JHAD1 cells. Increased autophagy continues 24 hours following C-PAC treatment as displayed in f), g), and h) with increased electron dense cargo supporting formation of degradative autophagic vacuoles. C-PAC treatment of OE19 cells induced cellular necrosis as illustrated in j) and k) by moderate chromatin clumping and nuclear disintegration or karyolysis (asterisk), accompanied by loss of plasma membrane integrity and marked loss of cellular contents as in k), l), and o) (extra-cytoplasmic cellular debris, red arrowhead). Cellular blebbing (k, n) and fragmentation of the nucleus (Karyorrhexis, black arrowhead) are evident. p) Autophagic vacuoles containing cellular debris indicate a low level of autophagy induction in OE19 cells. B. Representative photomicrographs (200X) of JHAD1 and OE33 EAC cells treated with C-PAC [50 μg/ml] for 6 and 24 hours, black arrows indicate apoptotic cells. The enlarged inset illustrates morphological changes of cytoplasmic swelling and vacuolization consistent with autophagy induction (as also noted in Figure 1D ). Monodansylcadaverine (MDC) staining and confocal microscopy were employed next to probe autophagic vacuoles in OE19, JHAD1 and OE33 cells following C-PAC treatment [50 μg/ml] for 24 hours. C-PAC treatment of OE33 cells resulted in increased accumulation of punctate staining as illustrated in large MDC positive vesicles (far right, bottom panel). C. To further investigate C-PAC induced autophagy, JHAD1, OE33 and OE19 cells were treated with C-PAC [50 μg/ml] or vehicle in triplicate and lysates collected at baseline and following 6, 24 and 48 hours of treatment; then, probed for LC3B, microtubule-associated protein 1 light chain 3 beta, an important early marker and effector of autophagy. Representative images are shown, expression values were normalized to the loading control GAPDH and a mean fold change from baseline or time of first detection calculated utilizing Quantity One software (Bio-Rad, Hercules, CA). Positive fold change values indicate increased expression and negative values reflect decreased expression. Figure 3C shows that C-PAC strongly ( P

    Techniques Used: Transmission Assay, Staining, Confocal Microscopy, Marker, Expressing, Software

    12) Product Images from "The Combination of Vitamin K3 and Vitamin C Has Synergic Activity against Forms of Trypanosoma cruzi through a Redox Imbalance Process"

    Article Title: The Combination of Vitamin K3 and Vitamin C Has Synergic Activity against Forms of Trypanosoma cruzi through a Redox Imbalance Process

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0144033

    Autophagic vacuoles in parasitic forms of Trypanosoma cruzi that were treated with vitamin K 3 and vitamin C, alone and combined, for 24 h using MDC labeling. (A) MDC fluorescence microscopy images: (a, a’) Untreated epimastigote forms. (b, b’) Epimastigote forms that were treated with 1.90 μM VK 3 . (c, c’) Epimastigote forms that were treated with 0.61 mM VC. (d, d’) Epimastigote forms that were treated with 1.90 μM VK 3 + 0.61 mM VC. (e, e’) Untreated trypomastigote forms. (f, f’) Trypomastigote forms that were treated with 0.35 μM VK 3 . (g, g’) Trypomastigote forms that were treated with 0.20 mM VC. (h, h’) Trypomastigote forms that were treated with 0.35 μM VK 3 + 0.20 mM VC. (i, i’) Untreated amastigote forms. (j, j’) Amastigote forms that were treated with 0.30 μM VK 3 . (k, k’) Amastigote forms that were treated with 0.18 mM VC. (l, l’) Amastigote forms that were treated with 0.30 μM VK 3 + 0.18 mM VC. Arrows, stained autophagic vacuoles; MDC, monodansylcadaverine; MDC + WTM: monodansylcadaverine + wortmannin. Scale bars: 20 μm. (B-D) MDC fluorescence obtained by ImageJ: (B) Epimastigote forms. (C) Trypomastigote forms. (D) Amastigote forms. * Indicate significant differences compared with the control group (untreated cells; p ≤ 0.05) and # indicate significant difference compared with the vitamins in combination without WTM ( p ≤ 0.05).
    Figure Legend Snippet: Autophagic vacuoles in parasitic forms of Trypanosoma cruzi that were treated with vitamin K 3 and vitamin C, alone and combined, for 24 h using MDC labeling. (A) MDC fluorescence microscopy images: (a, a’) Untreated epimastigote forms. (b, b’) Epimastigote forms that were treated with 1.90 μM VK 3 . (c, c’) Epimastigote forms that were treated with 0.61 mM VC. (d, d’) Epimastigote forms that were treated with 1.90 μM VK 3 + 0.61 mM VC. (e, e’) Untreated trypomastigote forms. (f, f’) Trypomastigote forms that were treated with 0.35 μM VK 3 . (g, g’) Trypomastigote forms that were treated with 0.20 mM VC. (h, h’) Trypomastigote forms that were treated with 0.35 μM VK 3 + 0.20 mM VC. (i, i’) Untreated amastigote forms. (j, j’) Amastigote forms that were treated with 0.30 μM VK 3 . (k, k’) Amastigote forms that were treated with 0.18 mM VC. (l, l’) Amastigote forms that were treated with 0.30 μM VK 3 + 0.18 mM VC. Arrows, stained autophagic vacuoles; MDC, monodansylcadaverine; MDC + WTM: monodansylcadaverine + wortmannin. Scale bars: 20 μm. (B-D) MDC fluorescence obtained by ImageJ: (B) Epimastigote forms. (C) Trypomastigote forms. (D) Amastigote forms. * Indicate significant differences compared with the control group (untreated cells; p ≤ 0.05) and # indicate significant difference compared with the vitamins in combination without WTM ( p ≤ 0.05).

    Techniques Used: Labeling, Fluorescence, Microscopy, Staining

    13) Product Images from "Polyphyllin VII Induces an Autophagic Cell Death by Activation of the JNK Pathway and Inhibition of PI3K/AKT/mTOR Pathway in HepG2 Cells"

    Article Title: Polyphyllin VII Induces an Autophagic Cell Death by Activation of the JNK Pathway and Inhibition of PI3K/AKT/mTOR Pathway in HepG2 Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0147405

    Induction of autophagy by PP7 in HepG2 cells. (A) Monodansylcadaverine (MDC) and (C) Acridine orange (AO) staining. HepG2 cells were treated with vehicle or PP7 for 24 h, stained with MDC and AO, and observed under InCell 2000 confocal microscope (20X objective). The mean fluorescence intensities of MDC (B) and AO (D) in the cells were calculated. (E) GFP-LC3-transfected HepG2 cells were incubated with or without PP7 for 24 h. The GFP-LC3 puncta was observed by InCell 2000 confocal microscope (60X objective). (F) Quantification of autophagic cells (E). Scale bars represent 20 μm. Western blot analysis of the LC3II, Beclin-1 and P62 in HepG2 cells treated with 1.32 μM PP7 for different times (G) or treated with varying concentrations of PP7 for 24 h (H). (I) and (J) were densitometic analysis of (G) and (H), respectively. Data are represented as means ± SD from 3 independent experiments. * P
    Figure Legend Snippet: Induction of autophagy by PP7 in HepG2 cells. (A) Monodansylcadaverine (MDC) and (C) Acridine orange (AO) staining. HepG2 cells were treated with vehicle or PP7 for 24 h, stained with MDC and AO, and observed under InCell 2000 confocal microscope (20X objective). The mean fluorescence intensities of MDC (B) and AO (D) in the cells were calculated. (E) GFP-LC3-transfected HepG2 cells were incubated with or without PP7 for 24 h. The GFP-LC3 puncta was observed by InCell 2000 confocal microscope (60X objective). (F) Quantification of autophagic cells (E). Scale bars represent 20 μm. Western blot analysis of the LC3II, Beclin-1 and P62 in HepG2 cells treated with 1.32 μM PP7 for different times (G) or treated with varying concentrations of PP7 for 24 h (H). (I) and (J) were densitometic analysis of (G) and (H), respectively. Data are represented as means ± SD from 3 independent experiments. * P

    Techniques Used: Staining, Microscopy, Fluorescence, Transfection, Incubation, Western Blot

    14) Product Images from "Lipopolysaccharide (LPS)-induced autophagy is involved in the restriction of Escherichia coli in peritoneal mesothelial cells"

    Article Title: Lipopolysaccharide (LPS)-induced autophagy is involved in the restriction of Escherichia coli in peritoneal mesothelial cells

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-13-255

    LPS stimulation induced autophagy in HMrSV5 cells. (A) Western blot analysis of Beclin-1 and LC3-II in HMrSV5 cells treated with LPS at various concentrations for 12 hours or 1 μg/ml LPS for the indicated time periods. β-actin was used as a loading control. (B) Densitometric anaysis of the blots showing the ratios of Beclin-1 and LC3-II to β-actin. (C) Transmission electron microscopy (TEM) of LPS-induced autophagy. Single-membrane phagosomes were seen in image 1. Image 2 shows typical double-membrane autophagosomes. Image 3 and 4 show multilayer structures. n, nucleus; av, autophagic vacuole; white arrows, single-membrane compartments; black arrows, double-membrane or multilayer structures. Scale bars: image1: 0.5 μm; image 2, 3 and 4: 200 nm . (D) Autophagic vacuoles were labeled with monodansylcadaverine (MDC, blue). Scale bars: 20 μm. (E) Graphs display quantitation of the number of autophagosomes per cross-sectioned cell (left panel) and the number of MDC-labeled autophagosomes per cell (right panel). Data are mean values ± SD (n ≥3). * p
    Figure Legend Snippet: LPS stimulation induced autophagy in HMrSV5 cells. (A) Western blot analysis of Beclin-1 and LC3-II in HMrSV5 cells treated with LPS at various concentrations for 12 hours or 1 μg/ml LPS for the indicated time periods. β-actin was used as a loading control. (B) Densitometric anaysis of the blots showing the ratios of Beclin-1 and LC3-II to β-actin. (C) Transmission electron microscopy (TEM) of LPS-induced autophagy. Single-membrane phagosomes were seen in image 1. Image 2 shows typical double-membrane autophagosomes. Image 3 and 4 show multilayer structures. n, nucleus; av, autophagic vacuole; white arrows, single-membrane compartments; black arrows, double-membrane or multilayer structures. Scale bars: image1: 0.5 μm; image 2, 3 and 4: 200 nm . (D) Autophagic vacuoles were labeled with monodansylcadaverine (MDC, blue). Scale bars: 20 μm. (E) Graphs display quantitation of the number of autophagosomes per cross-sectioned cell (left panel) and the number of MDC-labeled autophagosomes per cell (right panel). Data are mean values ± SD (n ≥3). * p

    Techniques Used: Western Blot, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Labeling, Quantitation Assay

    15) Product Images from "Levofolene modulates apoptosis induced by 5-fluorouracil through autophagy inhibition: Clinical and occupational implications"

    Article Title: Levofolene modulates apoptosis induced by 5-fluorouracil through autophagy inhibition: Clinical and occupational implications

    Journal: International Journal of Oncology

    doi: 10.3892/ijo.2015.2904

    Evaluation of autophagy in HaCaT cells treated with 5-fluorouracil (5-FU), 5-FU in combination with levofolene (LF) and doxorubicin (DOXO) after 24-48-72 h. (A) Flow cytometry overlay of monodansylcadaverine (MDC) fluorescence intensity. (B) Histogram of MDC mean fluorescence intensity (% of control). The bars represent means ± SD of three independent experiments. Asterisks indicate significant difference between 5-FU-treated vs. 5-FU+LF-treated cells ( ** P
    Figure Legend Snippet: Evaluation of autophagy in HaCaT cells treated with 5-fluorouracil (5-FU), 5-FU in combination with levofolene (LF) and doxorubicin (DOXO) after 24-48-72 h. (A) Flow cytometry overlay of monodansylcadaverine (MDC) fluorescence intensity. (B) Histogram of MDC mean fluorescence intensity (% of control). The bars represent means ± SD of three independent experiments. Asterisks indicate significant difference between 5-FU-treated vs. 5-FU+LF-treated cells ( ** P

    Techniques Used: Flow Cytometry, Cytometry, Fluorescence

    16) Product Images from "Transglutaminase down‐regulates the dimerization of epidermal growth factor receptor in rat perivenous and periportal hepatocytes"

    Article Title: Transglutaminase down‐regulates the dimerization of epidermal growth factor receptor in rat perivenous and periportal hepatocytes

    Journal: Cell Proliferation

    doi: 10.1111/j.1365-2184.2009.00622.x

    Effect of monodansylcadaverine (MDC) and retinoic acid (RA) on epidermal growth factor receptor (EGFR) dimerization induced by EGF treatment in primary cultured periportal hepatocytes (PPH) and perivenous hepatocytes (PVH). After EGF (10 −8 m) treatment at intervals 1 min, 5 min, 10 min, 20 min, 30 min, 1 h, 2 h, and 6 h, proteins were cross‐linked with bis‐(sulphosuccinimidyl) suberate (BS 3 ). Cell membranes obtained from hepatocytes were then lysed and protein samples were immunoblotted with anti‐EGFR antibodies. Band quantitation was performed with National Institutes of Health Image software. (a) Immunodetections of EGFRs (monomer and dimer) are shown. Results represent one typical experiment. Molecular weight markers are indicated on the left. (b) The histogram represents mean ± standard error of the mean of three independent experiments, expressed relative to the peak of EGFR dimer in PPH from MDC treated at 5 min following EGF treatment taken as 100%.
    Figure Legend Snippet: Effect of monodansylcadaverine (MDC) and retinoic acid (RA) on epidermal growth factor receptor (EGFR) dimerization induced by EGF treatment in primary cultured periportal hepatocytes (PPH) and perivenous hepatocytes (PVH). After EGF (10 −8 m) treatment at intervals 1 min, 5 min, 10 min, 20 min, 30 min, 1 h, 2 h, and 6 h, proteins were cross‐linked with bis‐(sulphosuccinimidyl) suberate (BS 3 ). Cell membranes obtained from hepatocytes were then lysed and protein samples were immunoblotted with anti‐EGFR antibodies. Band quantitation was performed with National Institutes of Health Image software. (a) Immunodetections of EGFRs (monomer and dimer) are shown. Results represent one typical experiment. Molecular weight markers are indicated on the left. (b) The histogram represents mean ± standard error of the mean of three independent experiments, expressed relative to the peak of EGFR dimer in PPH from MDC treated at 5 min following EGF treatment taken as 100%.

    Techniques Used: Cell Culture, Quantitation Assay, Software, Molecular Weight

    Effect of monodansylcadaverine (MDC) and retinoic acid (RA) on epidermal growth factor receptor (EGFR) phosphorylation induced by EGF treatment in primary cultured periportal hepatocytes (PPH) and perivenous hepatocytes (PVH). After EGF (10 −8 m) treatment for indicated times, cell membranes obtained from cultured hepatocytes were lysed and protein samples were immunoblotted with antibodies to EGFR phosphorylated at Y1173 and EGFR. To determine specific tyrosine phosphorylation of EGFR, PVDF membranes proved with anti‐phosphotyrosine antibody were stripped with a solution of 0.1 m glycine (pH 2.1) and reprobed with anti‐EGFR antibody. The band quantitation was performed with National Institutes of Health image software. (a) Immunodetections of EGFR phosphorylation and EGFR are shown. Results represent one typical experiment. (b) The histogram represents mean ± standard error of the mean of three independent experiments, expressed relative to the peak of the phosphorylation in EGFR in PPH from MDC treated at 5 min following EGF treatment taken as 100%.
    Figure Legend Snippet: Effect of monodansylcadaverine (MDC) and retinoic acid (RA) on epidermal growth factor receptor (EGFR) phosphorylation induced by EGF treatment in primary cultured periportal hepatocytes (PPH) and perivenous hepatocytes (PVH). After EGF (10 −8 m) treatment for indicated times, cell membranes obtained from cultured hepatocytes were lysed and protein samples were immunoblotted with antibodies to EGFR phosphorylated at Y1173 and EGFR. To determine specific tyrosine phosphorylation of EGFR, PVDF membranes proved with anti‐phosphotyrosine antibody were stripped with a solution of 0.1 m glycine (pH 2.1) and reprobed with anti‐EGFR antibody. The band quantitation was performed with National Institutes of Health image software. (a) Immunodetections of EGFR phosphorylation and EGFR are shown. Results represent one typical experiment. (b) The histogram represents mean ± standard error of the mean of three independent experiments, expressed relative to the peak of the phosphorylation in EGFR in PPH from MDC treated at 5 min following EGF treatment taken as 100%.

    Techniques Used: Cell Culture, Quantitation Assay, Software

    17) Product Images from "The Role of Deoxycytidine Kinase (dCK) in Radiation-Induced Cell Death"

    Article Title: The Role of Deoxycytidine Kinase (dCK) in Radiation-Induced Cell Death

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms17111939

    dCK promoted IR-induced autophagy in HeLa cells. ( A ) HeLa cells were treated with mock or IR (8 Gy). Staining of monodansylcadaverine (MDC) was detected by flow cytometry at 24 h after IR treatment; M1 represents fluorescent densities of MDC positive cells; ( B ) both dCK silencing cells and pSUPER cells were treated with 8 Gy radiation; 24 h later, staining of MDC was detected by flow cytometry; M1 represents fluorescent densities of MDC positive cells; ( C ) western blot analysis of microtubule-associated protein 1A/1B-light chain 3 (LC3) expression in the pSUPER and dCK knock down cells 24, 48 and 72 h after 8 Gy radiation. Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) was used as an internal standard; M1 represents fluorescent densities of MDC positive cells; ( D ) dCK knock-down HeLa cells were reintroduced with vector control, dCK wild-type, dCK-S74A mutation and dCK-S74E mutation. 36 h after transfection, cells were exposed to mock or IR (8 Gy). 24 h after IR, cells were harvested and subjected to western blot using the indicated antibodies.
    Figure Legend Snippet: dCK promoted IR-induced autophagy in HeLa cells. ( A ) HeLa cells were treated with mock or IR (8 Gy). Staining of monodansylcadaverine (MDC) was detected by flow cytometry at 24 h after IR treatment; M1 represents fluorescent densities of MDC positive cells; ( B ) both dCK silencing cells and pSUPER cells were treated with 8 Gy radiation; 24 h later, staining of MDC was detected by flow cytometry; M1 represents fluorescent densities of MDC positive cells; ( C ) western blot analysis of microtubule-associated protein 1A/1B-light chain 3 (LC3) expression in the pSUPER and dCK knock down cells 24, 48 and 72 h after 8 Gy radiation. Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) was used as an internal standard; M1 represents fluorescent densities of MDC positive cells; ( D ) dCK knock-down HeLa cells were reintroduced with vector control, dCK wild-type, dCK-S74A mutation and dCK-S74E mutation. 36 h after transfection, cells were exposed to mock or IR (8 Gy). 24 h after IR, cells were harvested and subjected to western blot using the indicated antibodies.

    Techniques Used: Staining, Flow Cytometry, Cytometry, Western Blot, Expressing, Plasmid Preparation, Mutagenesis, Transfection

    18) Product Images from "Antitumor effect of matrine in human hepatoma G2 cells by inducing apoptosis and autophagy"

    Article Title: Antitumor effect of matrine in human hepatoma G2 cells by inducing apoptosis and autophagy

    Journal: World Journal of Gastroenterology : WJG

    doi: 10.3748/wjg.v16.i34.4281

    Monodansylcadaverine-labeled vacuoles in HepG2 cells. Autophagic vacuoles were labeled with 0.05 mmol/L monodansylcadaverine (MDC) in phosphate-buffered saline (PBS) at 37°C for 10 min. The fluorescent density and the MDC-labeled particles in
    Figure Legend Snippet: Monodansylcadaverine-labeled vacuoles in HepG2 cells. Autophagic vacuoles were labeled with 0.05 mmol/L monodansylcadaverine (MDC) in phosphate-buffered saline (PBS) at 37°C for 10 min. The fluorescent density and the MDC-labeled particles in

    Techniques Used: Labeling

    19) Product Images from "Radiosensitivity of human ovarian cancer cells is enhanced by pseudolaric acid B due to the inhibition of the Ras/Raf/ERK signaling pathway"

    Article Title: Radiosensitivity of human ovarian cancer cells is enhanced by pseudolaric acid B due to the inhibition of the Ras/Raf/ERK signaling pathway

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.5500

    Autophagy is not affected by PAB in SKOV-3 cells. SKOV-3 cells were exposed to a range of PAB concentrations and 4 Gy radiation. After 24 h, (A) cells underwent MDC staining and the results were measured by flow cytometry or (B) the expression of LC3-I, LC3-II and ATG5 were analyzed by western blot analysis. PAB, pseudolaric acid B; MDC, monodansylcadaverine; LC3-I, microtubule-associated protein 1 light chain 3; LC3-II, microtubule-associated protein 2 light chain 3; ATG5, autophagy protein 5.
    Figure Legend Snippet: Autophagy is not affected by PAB in SKOV-3 cells. SKOV-3 cells were exposed to a range of PAB concentrations and 4 Gy radiation. After 24 h, (A) cells underwent MDC staining and the results were measured by flow cytometry or (B) the expression of LC3-I, LC3-II and ATG5 were analyzed by western blot analysis. PAB, pseudolaric acid B; MDC, monodansylcadaverine; LC3-I, microtubule-associated protein 1 light chain 3; LC3-II, microtubule-associated protein 2 light chain 3; ATG5, autophagy protein 5.

    Techniques Used: Staining, Flow Cytometry, Cytometry, Expressing, Western Blot

    20) Product Images from "Autophagy in human articular chondrocytes is cytoprotective following glucocorticoid stimulation"

    Article Title: Autophagy in human articular chondrocytes is cytoprotective following glucocorticoid stimulation

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2014.2102

    State of autophagy in human chondrocytes following Dex treatment. (A) MDC-labeled vacuoles were examined by fluorescence microscopy (magnification, ×400). (B) Formation of autophagic vacuoles were observed by TEM (magnification, ×15,000). Autophagic vacuoles are indicated by black arrows. (C) Western blot analysis of LC3-I, LC3-II and beclin-1 following incubation with Dex for different times. β-actin was used as a internal control. (D) The specific activity index (%) was obtained by flow cytometry. (E) Levels of proteins were normalized with respect to the β-actin band density using Bio-Rad Quantity One software. Data are expressed as the mean ± standard deviation. Dex, dexamethasone; MDC, monodansylcadaverine; TEM, transmission electron microscopy; LC3, microtubule-associated protein light chain 3.
    Figure Legend Snippet: State of autophagy in human chondrocytes following Dex treatment. (A) MDC-labeled vacuoles were examined by fluorescence microscopy (magnification, ×400). (B) Formation of autophagic vacuoles were observed by TEM (magnification, ×15,000). Autophagic vacuoles are indicated by black arrows. (C) Western blot analysis of LC3-I, LC3-II and beclin-1 following incubation with Dex for different times. β-actin was used as a internal control. (D) The specific activity index (%) was obtained by flow cytometry. (E) Levels of proteins were normalized with respect to the β-actin band density using Bio-Rad Quantity One software. Data are expressed as the mean ± standard deviation. Dex, dexamethasone; MDC, monodansylcadaverine; TEM, transmission electron microscopy; LC3, microtubule-associated protein light chain 3.

    Techniques Used: Labeling, Fluorescence, Microscopy, Transmission Electron Microscopy, Western Blot, Incubation, Activity Assay, Flow Cytometry, Cytometry, Software, Standard Deviation, Transmission Assay, Electron Microscopy

    21) Product Images from "PTEN Activation by DNA Damage Induces Protective Autophagy in Response to Cucurbitacin B in Hepatocellular Carcinoma Cells"

    Article Title: PTEN Activation by DNA Damage Induces Protective Autophagy in Response to Cucurbitacin B in Hepatocellular Carcinoma Cells

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2016/4313204

    Cuc B induced protective autophagy. Cells were treated with Cuc B for 6 h and stained with MDC (20x) (a) and the protein expression was detected by Western blot (b and d). Cells were treated with Cuc B (100 nM) and the expression of LC3 was detected by Western blot (c). Cells were pretreated with 3-MA (2.5 mM) (e) or CQ (10 μ M) (f) for 2 h and then cotreated with Cuc B for 6 h, and the cell viability was determined by MTT. Cuc B, Cucurbitacin B; MDC, monodansylcadaverine. ∗∗ p
    Figure Legend Snippet: Cuc B induced protective autophagy. Cells were treated with Cuc B for 6 h and stained with MDC (20x) (a) and the protein expression was detected by Western blot (b and d). Cells were treated with Cuc B (100 nM) and the expression of LC3 was detected by Western blot (c). Cells were pretreated with 3-MA (2.5 mM) (e) or CQ (10 μ M) (f) for 2 h and then cotreated with Cuc B for 6 h, and the cell viability was determined by MTT. Cuc B, Cucurbitacin B; MDC, monodansylcadaverine. ∗∗ p

    Techniques Used: Staining, Expressing, Western Blot, MTT Assay

    22) Product Images from "Targeted inhibition of mammalian target of rapamycin (mTOR) enhances radiosensitivity in pancreatic carcinoma cells"

    Article Title: Targeted inhibition of mammalian target of rapamycin (mTOR) enhances radiosensitivity in pancreatic carcinoma cells

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S42390

    MDC-labeled autophagic vacuoles in PC-2 cells by fluorescence microscope after RPM treatment. Autophagic vacuoles were labeled with 0.05 mmol/L MDC in PBS, at 37°C for 10 minutes. ( A ) 0 nmol/L RPM group; ( B ) 10 nmol/L RPM group; and ( C ) 15 nmol/L RPM group (×200). Abbreviations: MDC, monodansylcadaverine; PBS, phosphate-buffered saline; RPM, rapamycin.
    Figure Legend Snippet: MDC-labeled autophagic vacuoles in PC-2 cells by fluorescence microscope after RPM treatment. Autophagic vacuoles were labeled with 0.05 mmol/L MDC in PBS, at 37°C for 10 minutes. ( A ) 0 nmol/L RPM group; ( B ) 10 nmol/L RPM group; and ( C ) 15 nmol/L RPM group (×200). Abbreviations: MDC, monodansylcadaverine; PBS, phosphate-buffered saline; RPM, rapamycin.

    Techniques Used: Labeling, Fluorescence, Microscopy

    23) Product Images from "VEGF-A selectively inhibits FLT1 ectodomain shedding independent of receptor activation and receptor endocytosis"

    Article Title: VEGF-A selectively inhibits FLT1 ectodomain shedding independent of receptor activation and receptor endocytosis

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00247.2017

    Effect of N-acetylleucylleucylnorleucinal (ALLN), bafilomycin, and monodansylcadaverine (MDC) on the abundance and cleavage of Flt1 and FLT1ΔCTD. A : HEK293 cells transfected with hemagglutinin (HA)-Flt1-Myc-Flag (Flt1) or HA-FLT1ΔCTD (deleted COOH-terminal domain, ΔCTD) and incubated with ALLN (10 µM) and/or 100 ng/ml VEGF-A for 24 h and then immunoblotted for Flag, HA, heat shock protein 90 (HSP90), and epidermal growth factor receptor (EGFR). COOH-terminal fragments (CTF1 and CTF2) and the NH 2 -terminal fragment (NTF) are seen in membrane preps, cytosol, and conditioned media (CM), respectively. EGFR and HSP90 are used as loading controls. ALLN does not reverse the VEGF-A inhibition of FLT1 cleavage. B : HEK293 cells transfected with Flt1 or ΔCTD and incubated with 250 nM bafilomycin A (BafA) for 8 h and/or 100 ng/ml VEGF-A for 7.5 h and then immunoblotted for Flag, HA, and tubulin. Full-length Flt1 (FL) and the NTF are seen in lysates and CM, respectively. BafA does not reverse the inhibition seen with VEGF-A. C : HEK293 cells transfected with HA-Flt1-Myc-Flag ( A ) or FLT1ΔCTD ( B ) and incubated with 125 µM MDC and/or 100 ng/ml VEGF-A for 24 h and then immunoblotted for Flag, HA, and tubulin. MDC does not reverse the inhibition seen with VEGF-A. D and E : AG1-G1-FLT1 cells treated with BafA or MDC with or without 100 ng/ml VEGF-A for 4 h and then immunoblotted with AF321 and SC-316 antibodies to detect NTF and FL protein of FLT1 respectively and with HSP90 as a loading control. BafA and MDC do not reverse the inhibition seen with VEGF-A. A representative immunoblot is seen in D . The effect of BafA was replicated in additional experiments and the pooled NTF data quantified by densitometry are shown in E . Means ± SD, n = 3. * P
    Figure Legend Snippet: Effect of N-acetylleucylleucylnorleucinal (ALLN), bafilomycin, and monodansylcadaverine (MDC) on the abundance and cleavage of Flt1 and FLT1ΔCTD. A : HEK293 cells transfected with hemagglutinin (HA)-Flt1-Myc-Flag (Flt1) or HA-FLT1ΔCTD (deleted COOH-terminal domain, ΔCTD) and incubated with ALLN (10 µM) and/or 100 ng/ml VEGF-A for 24 h and then immunoblotted for Flag, HA, heat shock protein 90 (HSP90), and epidermal growth factor receptor (EGFR). COOH-terminal fragments (CTF1 and CTF2) and the NH 2 -terminal fragment (NTF) are seen in membrane preps, cytosol, and conditioned media (CM), respectively. EGFR and HSP90 are used as loading controls. ALLN does not reverse the VEGF-A inhibition of FLT1 cleavage. B : HEK293 cells transfected with Flt1 or ΔCTD and incubated with 250 nM bafilomycin A (BafA) for 8 h and/or 100 ng/ml VEGF-A for 7.5 h and then immunoblotted for Flag, HA, and tubulin. Full-length Flt1 (FL) and the NTF are seen in lysates and CM, respectively. BafA does not reverse the inhibition seen with VEGF-A. C : HEK293 cells transfected with HA-Flt1-Myc-Flag ( A ) or FLT1ΔCTD ( B ) and incubated with 125 µM MDC and/or 100 ng/ml VEGF-A for 24 h and then immunoblotted for Flag, HA, and tubulin. MDC does not reverse the inhibition seen with VEGF-A. D and E : AG1-G1-FLT1 cells treated with BafA or MDC with or without 100 ng/ml VEGF-A for 4 h and then immunoblotted with AF321 and SC-316 antibodies to detect NTF and FL protein of FLT1 respectively and with HSP90 as a loading control. BafA and MDC do not reverse the inhibition seen with VEGF-A. A representative immunoblot is seen in D . The effect of BafA was replicated in additional experiments and the pooled NTF data quantified by densitometry are shown in E . Means ± SD, n = 3. * P

    Techniques Used: Transfection, Incubation, Inhibition

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    Article Snippet: .. Briefly, RAECMs were incubated with methyl-β-cyclodextrin (10–200 μM; Sigma), monodansylcadaverine (200 μM; Sigma), chlorpromazine (28 μM; Sigma), latrunculin B (20 μM; Sigma), cyctochalasin D (10 μM; Sigma), or dynasore (80 μM; Sigma) in both apical and basolateral fluids for 30 minutes. ..

    other:

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    Labeling:

    Article Title: Caveolin-1 mediated uptake via langerin restricts HIV-1 infection in human Langerhans cells
    Article Snippet: .. Antibodies and reagents The following antibodies were used: Rabbit-anti-Caveolin-1 (Cell Signalling); Goat-anti-langerin (R & D); DCGM4-PE (mouse-anti-langerin; Beckman Coulter); KC57-RD1-PE (mouse-anti-p24; Beckman Coulter); mouse-anti-CD1a-FITC (BD Pharmingen); anti-LAMP2 (Abcam); anti-CD63 (BD); sheep-anti-p24 (Aalto); 10E2 (anti-langerin [ ]); 10E2 coupled to Alexa-647 (Alexa-647 labeling kit); Streptavidin-Alexa-488; Goat-anti-Mouse IgG1 Alexa 546; Goat-anti-Rabbit Alexa 488 (5 μg/ml, all Invitrogen); 15 nm protein A-gold; 10 nm protein G-gold (both Aurion); prot A/G plus agarose beads (Santa Cruz); filipin complex (Sigma Aldrich); Dispase II (Roche Diagnostics); monodansylcadaverine (Sigma Aldrich). ..

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    Millipore monodansylcadaverine mdc
    <t>Monodansylcadaverine</t> <t>(MDC)</t> staining of cytoplasmic vacuoles after propofol treatment in human keratinocytes. Significant accumulation of autophagy-specific staining of MDC was observed around the nuclei in PPC/H 2 O 2 group cells.
    Monodansylcadaverine Mdc, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Monodansylcadaverine (MDC) staining of cytoplasmic vacuoles after propofol treatment in human keratinocytes. Significant accumulation of autophagy-specific staining of MDC was observed around the nuclei in PPC/H 2 O 2 group cells.

    Journal: Journal of Dental Anesthesia and Pain Medicine

    Article Title: Propofol protects human keratinocytes from oxidative stress via autophagy expression

    doi: 10.17245/jdapm.2017.17.1.21

    Figure Lengend Snippet: Monodansylcadaverine (MDC) staining of cytoplasmic vacuoles after propofol treatment in human keratinocytes. Significant accumulation of autophagy-specific staining of MDC was observed around the nuclei in PPC/H 2 O 2 group cells.

    Article Snippet: The following reagents were obtained commercially: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), acridine orange (AO), monodansylcadaverine (MDC), and 3-methyladenine (3-MA, class III PI3K inhibitor) (Calbiochem, La Jolla, CA, USA).

    Techniques: Staining

    Analyses of the role of endocytosis routes and paracellular permeability in antibody transfer to the draining LNs. ( A ) Confocal and flow cytometric analyses of draining LNs of WT and Cav1 −/− mice after s.c. administration of the indicated fluorochrome-conjugated antibodies (1 μg, t = 5 min, n = 4). The cells were stained ex vivo for CD3 for flow cytometric analysis. Scale bars: 20 μm. ( B ) Experimental outline for topical application of endocytosis inhibitors (and controls) to surgically exposed LNs. ( C – G ) Analyses of transcytosis of the indicated s.c. administered fluorochrome-conjugated antibodies (5 μg each, t = 5 min) after topical pretreatment of the draining LNs with ( C – F ) endocytosis inhibitors or ( G ) a LEC junctional stabilizer. ( C ) Pitstop 2 (a clathrin inhibitor) and a Pitstop 2 negative control; ( D ) monodansylcadaverine (MDC) (a clathrin inhibitor) and vehicle; ( E ) EIPA (a macropinocytosis inhibitor) and vehicle; ( F ) imipramine (a macropinocytosis inhibitor) and vehicle; and ( G ) adrenomedullin and vehicle. The cells were stained ex vivo for CD3. In the bar graphs, each dot represents 1 LN, and data are the mean ± SD. Statistical significance was determined by Mann-Whitney U test.

    Journal: The Journal of Clinical Investigation

    Article Title: Transcytosis route mediates rapid delivery of intact antibodies to draining lymph nodes

    doi: 10.1172/JCI125740

    Figure Lengend Snippet: Analyses of the role of endocytosis routes and paracellular permeability in antibody transfer to the draining LNs. ( A ) Confocal and flow cytometric analyses of draining LNs of WT and Cav1 −/− mice after s.c. administration of the indicated fluorochrome-conjugated antibodies (1 μg, t = 5 min, n = 4). The cells were stained ex vivo for CD3 for flow cytometric analysis. Scale bars: 20 μm. ( B ) Experimental outline for topical application of endocytosis inhibitors (and controls) to surgically exposed LNs. ( C – G ) Analyses of transcytosis of the indicated s.c. administered fluorochrome-conjugated antibodies (5 μg each, t = 5 min) after topical pretreatment of the draining LNs with ( C – F ) endocytosis inhibitors or ( G ) a LEC junctional stabilizer. ( C ) Pitstop 2 (a clathrin inhibitor) and a Pitstop 2 negative control; ( D ) monodansylcadaverine (MDC) (a clathrin inhibitor) and vehicle; ( E ) EIPA (a macropinocytosis inhibitor) and vehicle; ( F ) imipramine (a macropinocytosis inhibitor) and vehicle; and ( G ) adrenomedullin and vehicle. The cells were stained ex vivo for CD3. In the bar graphs, each dot represents 1 LN, and data are the mean ± SD. Statistical significance was determined by Mann-Whitney U test.

    Article Snippet: The following endocytosis inhibitors and controls were used: clathrin inhibitor Pitstop 2 ( ) (Abcam) and its specific negative control (Pitstop 2 control, Abcam) both at 75 μM; the clathrin inhibitor monodansylcadaverine ( ) (MilliporeSigma; 100 μM) and its DMSO vehicle control; the macropinocytosis inhibitor EIPA ( ) [5-( N -ethyl- N -isopropyl amiloride, MilliporeSigma; 100 μM] and its DMSO vehicle control; the macropinocytosis inhibitor imipramine ( ) (MilliporeSigma, 15 μM) and its PBS vehicle control; the dynamin inhibitor Dyngo-4a ( ) (MilliporeSigma, 400 μM) and its DMSO vehicle control; and the dynamin inhibitor Dynole 34-2 (39 (Abcam) and its specific control Dynole 31-2 (Abcam), both at 75 μM.

    Techniques: Permeability, Flow Cytometry, Mouse Assay, Staining, Ex Vivo, Negative Control, MANN-WHITNEY