Structured Review

National Centre for Cell Science a549 cell lines
Confocal microscopic images of <t>A549</t> cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.
A549 Cell Lines, supplied by National Centre for Cell Science, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/a549 cell lines/product/National Centre for Cell Science
Average 92 stars, based on 2 article reviews
Price from $9.99 to $1999.99
a549 cell lines - by Bioz Stars, 2020-09
92/100 stars

Images

1) Product Images from "Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies"

Article Title: Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies

Journal: ACS Omega

doi: 10.1021/acsomega.8b01014

Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.
Figure Legend Snippet: Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.

Techniques Used: Staining

FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.
Figure Legend Snippet: FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.

Techniques Used: FACS, Expressing

Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.
Figure Legend Snippet: Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.

Techniques Used: Transfection

2) Product Images from "Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies"

Article Title: Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies

Journal: ACS Omega

doi: 10.1021/acsomega.8b01014

Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.
Figure Legend Snippet: Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.

Techniques Used: Staining

FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.
Figure Legend Snippet: FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.

Techniques Used: FACS, Expressing

Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.
Figure Legend Snippet: Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.

Techniques Used: Transfection

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Article Title: An Adaptogen: Withaferin A Ameliorates in Vitro and in Vivo Pulmonary Fibrosis by Modulating the Interplay of Fibrotic, Matricelluar Proteins, and Cytokines
Article Snippet: .. Cell Culture HFL1 cells were procured from ATCC (ATCC® CCL153TM ) and A549 cells were purchased from National Centre for Cell Science (NCCS, Pune, India). .. HFL1 and A549 cells were cultured in F-12K medium (ATCC) and RPMI medium (Sigma-Aldrich, United States) respectively; supplemented with 10% fetal bovine serum and 1% anti-biotic solution (Invitrogen, United States).

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    National Centre for Cell Science lung adenocarcinoma cell line a549
    Percent <t>A549</t> cell viabilities after 72 h for various treatments
    Lung Adenocarcinoma Cell Line A549, supplied by National Centre for Cell Science, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lung adenocarcinoma cell line a549/product/National Centre for Cell Science
    Average 88 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    lung adenocarcinoma cell line a549 - by Bioz Stars, 2020-09
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    National Centre for Cell Science a549
    Capsaicin suppresses VEGF expression by SMAR1-mediated down regulation of Cox-2. (A) Control-/SMAR1-shRNA ( left panel ) or control-vector/SMAR1-cDNA ( right panel ) transfected <t>Hy-A549</t> cells were treated with capsaicin were assayed for Cox-2 expression by Western blot and RT-PCR. Transfection efficiency was verified by Western blot ( bottom panels ). (B) Cox-2 promoter activity was checked in Hy-A549. Schematic representation of the Cox-2 promoter showing the probable SMAR1-binding sites predicted by using the MARWIZ software, PCR run with eight different sets of primers designed for each probable MAR binding site spreading over four regions (−141 bp to −421 bp; −631 bp to −1121 bp; −1262 bp to −1331 bp; −1471 bp to −1891 bp). ChIP assay with anti-SMAR1 was performed on MAR-binding regions of Cox-2 promoter. Input and control IgG was used as internal control and negative control. (C) The relative abundance of SMAR1 on Cox-2 promoter was analyzed in control and capsaicin treated Hy-A549 cells at binding sites 6 7 after ChIP of Cox-2 promoter with anti-SMAR1( left panel ) and represented graphically ( right panel ). (D) Control-/SMAR1-shRNA-transfected Hy-A549 cells were treated with capsaicin and analyzed for reporter HIF-1α and VEGF gene expression by RT-PCR. α-Actin/GAPDH were used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.
    A549, supplied by National Centre for Cell Science, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a549/product/National Centre for Cell Science
    Average 92 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    a549 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    Image Search Results


    Percent A549 cell viabilities after 72 h for various treatments

    Journal: Cancer Nanotechnology

    Article Title: PLGA nanoparticles loaded with etoposide and quercetin dihydrate individually: in vitro cell line study to ensure advantage of combination therapy

    doi: 10.1007/s12645-012-0027-y

    Figure Lengend Snippet: Percent A549 cell viabilities after 72 h for various treatments

    Article Snippet: Lung adenocarcinoma cell line A549 was purchased from National Centre for Cell Science (NCCS), Pune, India. (MTT) 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide was purchased from HiMedia, India.

    Techniques:

    Percent A549 cell viabilities after 24 h for various treatments

    Journal: Cancer Nanotechnology

    Article Title: PLGA nanoparticles loaded with etoposide and quercetin dihydrate individually: in vitro cell line study to ensure advantage of combination therapy

    doi: 10.1007/s12645-012-0027-y

    Figure Lengend Snippet: Percent A549 cell viabilities after 24 h for various treatments

    Article Snippet: Lung adenocarcinoma cell line A549 was purchased from National Centre for Cell Science (NCCS), Pune, India. (MTT) 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide was purchased from HiMedia, India.

    Techniques:

    Percent A549 cell viabilities after 48 h for various treatments

    Journal: Cancer Nanotechnology

    Article Title: PLGA nanoparticles loaded with etoposide and quercetin dihydrate individually: in vitro cell line study to ensure advantage of combination therapy

    doi: 10.1007/s12645-012-0027-y

    Figure Lengend Snippet: Percent A549 cell viabilities after 48 h for various treatments

    Article Snippet: Lung adenocarcinoma cell line A549 was purchased from National Centre for Cell Science (NCCS), Pune, India. (MTT) 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide was purchased from HiMedia, India.

    Techniques:

    Capsaicin suppresses VEGF expression by SMAR1-mediated down regulation of Cox-2. (A) Control-/SMAR1-shRNA ( left panel ) or control-vector/SMAR1-cDNA ( right panel ) transfected Hy-A549 cells were treated with capsaicin were assayed for Cox-2 expression by Western blot and RT-PCR. Transfection efficiency was verified by Western blot ( bottom panels ). (B) Cox-2 promoter activity was checked in Hy-A549. Schematic representation of the Cox-2 promoter showing the probable SMAR1-binding sites predicted by using the MARWIZ software, PCR run with eight different sets of primers designed for each probable MAR binding site spreading over four regions (−141 bp to −421 bp; −631 bp to −1121 bp; −1262 bp to −1331 bp; −1471 bp to −1891 bp). ChIP assay with anti-SMAR1 was performed on MAR-binding regions of Cox-2 promoter. Input and control IgG was used as internal control and negative control. (C) The relative abundance of SMAR1 on Cox-2 promoter was analyzed in control and capsaicin treated Hy-A549 cells at binding sites 6 7 after ChIP of Cox-2 promoter with anti-SMAR1( left panel ) and represented graphically ( right panel ). (D) Control-/SMAR1-shRNA-transfected Hy-A549 cells were treated with capsaicin and analyzed for reporter HIF-1α and VEGF gene expression by RT-PCR. α-Actin/GAPDH were used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Journal: PLoS ONE

    Article Title: Capsaicin-Induced Activation of p53-SMAR1 Auto-Regulatory Loop Down-Regulates VEGF in Non-Small Cell Lung Cancer to Restrain Angiogenesis

    doi: 10.1371/journal.pone.0099743

    Figure Lengend Snippet: Capsaicin suppresses VEGF expression by SMAR1-mediated down regulation of Cox-2. (A) Control-/SMAR1-shRNA ( left panel ) or control-vector/SMAR1-cDNA ( right panel ) transfected Hy-A549 cells were treated with capsaicin were assayed for Cox-2 expression by Western blot and RT-PCR. Transfection efficiency was verified by Western blot ( bottom panels ). (B) Cox-2 promoter activity was checked in Hy-A549. Schematic representation of the Cox-2 promoter showing the probable SMAR1-binding sites predicted by using the MARWIZ software, PCR run with eight different sets of primers designed for each probable MAR binding site spreading over four regions (−141 bp to −421 bp; −631 bp to −1121 bp; −1262 bp to −1331 bp; −1471 bp to −1891 bp). ChIP assay with anti-SMAR1 was performed on MAR-binding regions of Cox-2 promoter. Input and control IgG was used as internal control and negative control. (C) The relative abundance of SMAR1 on Cox-2 promoter was analyzed in control and capsaicin treated Hy-A549 cells at binding sites 6 7 after ChIP of Cox-2 promoter with anti-SMAR1( left panel ) and represented graphically ( right panel ). (D) Control-/SMAR1-shRNA-transfected Hy-A549 cells were treated with capsaicin and analyzed for reporter HIF-1α and VEGF gene expression by RT-PCR. α-Actin/GAPDH were used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Article Snippet: Cell culture Human cancer cell lines, A549, MCF-7, HeLa, HCT-15, and normal lung fibroblast WI-38, were obtained from National Centre for Cell Science, India, and maintained at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS (Lonza, NH), L-glutamine (2 mM), sodium pyruvate (100 µg/ml), nonessential amino acids (100 µM), streptomycin (100 µg/ml), penicillin (50 U/ml; Invitogen, CA) .

    Techniques: Expressing, shRNA, Plasmid Preparation, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Binding Assay, Software, Polymerase Chain Reaction, Chromatin Immunoprecipitation, Negative Control

    Capsaicin selectively inhibits VEGF secretion to retard Hy-A549 cell-induced HUVEC cell migration. (A) Representative phase contrast photomicrographs demonstrating HUVEC migration upon incubation with spent media of Brefeldin-A-pretreated, capsaicin (37.5 µM)-treated Hy-A549 cells ( left panel ). Percent cell migrated in the wound area has been represented graphically ( right panel ). (B) Immunoblots showing expression profiles of pro-angiogenic factors VEGF, bFGF, EGF, TGF-β, in presence or absence of capsaicin. (C) Secreted VEGF from cell-free supernatant of Hy-A549 was quantified by ELISA assay ( left panel ). Time-dependent expression profiles of VEGF-mRNA/-protein in capsaicin-treated Hy-A549 cells were determined by Western blot and RT-PCR respectively ( middle panel ). Capsaicin-treated Hy-A549 cells were examined for time-dependent variation in the expression profiles of VEGF by quantitative real time PCR analysis and represented graphically ( right panel ). (D) Immuno-fluorescent images (60x magnification) showing time-dependent pattern of VEGF protein (TRITC-fluorescent) in capsaicin-treated Hy-A549 cells were represented along with nuclear staining (DAPI: blue). (E) Representative images of HUVEC migration upon incubation with (i) recombinant VEGF-supplemented control media, or VEGF-supplemented spent media of capsaicin-treated Hy-A549 cells, or with (ii) anti-VEGF-treated Hy-A549 spent media ( left panel ). Percent cell migrated in the wound area is being represented graphically ( right panel ). (F) Representative images of capillary-like sprout formation by HUVECs upon incubation with recombinant VEGF-supplemented spent media of capsaicin-treated Hy-A549 cells or with anti-VEGF-treated Hy-A549 spent media. GAPDH/α-Actin was used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Journal: PLoS ONE

    Article Title: Capsaicin-Induced Activation of p53-SMAR1 Auto-Regulatory Loop Down-Regulates VEGF in Non-Small Cell Lung Cancer to Restrain Angiogenesis

    doi: 10.1371/journal.pone.0099743

    Figure Lengend Snippet: Capsaicin selectively inhibits VEGF secretion to retard Hy-A549 cell-induced HUVEC cell migration. (A) Representative phase contrast photomicrographs demonstrating HUVEC migration upon incubation with spent media of Brefeldin-A-pretreated, capsaicin (37.5 µM)-treated Hy-A549 cells ( left panel ). Percent cell migrated in the wound area has been represented graphically ( right panel ). (B) Immunoblots showing expression profiles of pro-angiogenic factors VEGF, bFGF, EGF, TGF-β, in presence or absence of capsaicin. (C) Secreted VEGF from cell-free supernatant of Hy-A549 was quantified by ELISA assay ( left panel ). Time-dependent expression profiles of VEGF-mRNA/-protein in capsaicin-treated Hy-A549 cells were determined by Western blot and RT-PCR respectively ( middle panel ). Capsaicin-treated Hy-A549 cells were examined for time-dependent variation in the expression profiles of VEGF by quantitative real time PCR analysis and represented graphically ( right panel ). (D) Immuno-fluorescent images (60x magnification) showing time-dependent pattern of VEGF protein (TRITC-fluorescent) in capsaicin-treated Hy-A549 cells were represented along with nuclear staining (DAPI: blue). (E) Representative images of HUVEC migration upon incubation with (i) recombinant VEGF-supplemented control media, or VEGF-supplemented spent media of capsaicin-treated Hy-A549 cells, or with (ii) anti-VEGF-treated Hy-A549 spent media ( left panel ). Percent cell migrated in the wound area is being represented graphically ( right panel ). (F) Representative images of capillary-like sprout formation by HUVECs upon incubation with recombinant VEGF-supplemented spent media of capsaicin-treated Hy-A549 cells or with anti-VEGF-treated Hy-A549 spent media. GAPDH/α-Actin was used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Article Snippet: Cell culture Human cancer cell lines, A549, MCF-7, HeLa, HCT-15, and normal lung fibroblast WI-38, were obtained from National Centre for Cell Science, India, and maintained at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS (Lonza, NH), L-glutamine (2 mM), sodium pyruvate (100 µg/ml), nonessential amino acids (100 µM), streptomycin (100 µg/ml), penicillin (50 U/ml; Invitogen, CA) .

    Techniques: Migration, Incubation, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Staining, Recombinant

    Effect of capsaicin on lung cancer cell spent medium-induced endothelial cell migration and network formation. (A) Migration of HUVECs in presence or absence of spent media of NME, WI-38, A549 and Hy-A549 (CoCl 2 -stimulated to mimic hypoxic condition required for tumor-induced angiogenesis) were subjected to bi-directional wound healing assay for 24 h ( left panel ). The number of cells migrated in the wound area are represented graphically ( right panel ). (B) Representative images of HUVEC migration upon incubation with capsaicin-treated (0–50 µM) Hy-A549 cell spent medium ( left panel ). Percent cell migrated in the wound area has been represented graphically ( right panel ). (C) Hy-A549 cells were treated with capsaicin in a dose-dependent manner for 24 h and cell viability was scored by trypan blue dye-exclusion assay ( left panel ). Hy-A549 cells, treated with capsaicin (37.5 µM), were subjected to Annexin-V-FITC/PI binding and analyzed flow cytometrically for the determination of percent early apoptosis ( right panel ). (D) Cytotoxic effect of different doses of capsaisin on HUVEC cells were measured by trypan blue dye-exclusion assay. (E) Graphical representation of HUVEC migration upon incubation with spent media from capsaicin-treated (37.5 µM) HBL-100, HCT-15, HeLa and A549. (F) Representative images of capillary-like sprout formation by HUVECs in presence of media alone or spent media of WI-38/A549/Hy-A549/capsaicin-treated Hy-A549. Values are mean ± SEM of three independent experiments in each case or representative of typical experiment.

    Journal: PLoS ONE

    Article Title: Capsaicin-Induced Activation of p53-SMAR1 Auto-Regulatory Loop Down-Regulates VEGF in Non-Small Cell Lung Cancer to Restrain Angiogenesis

    doi: 10.1371/journal.pone.0099743

    Figure Lengend Snippet: Effect of capsaicin on lung cancer cell spent medium-induced endothelial cell migration and network formation. (A) Migration of HUVECs in presence or absence of spent media of NME, WI-38, A549 and Hy-A549 (CoCl 2 -stimulated to mimic hypoxic condition required for tumor-induced angiogenesis) were subjected to bi-directional wound healing assay for 24 h ( left panel ). The number of cells migrated in the wound area are represented graphically ( right panel ). (B) Representative images of HUVEC migration upon incubation with capsaicin-treated (0–50 µM) Hy-A549 cell spent medium ( left panel ). Percent cell migrated in the wound area has been represented graphically ( right panel ). (C) Hy-A549 cells were treated with capsaicin in a dose-dependent manner for 24 h and cell viability was scored by trypan blue dye-exclusion assay ( left panel ). Hy-A549 cells, treated with capsaicin (37.5 µM), were subjected to Annexin-V-FITC/PI binding and analyzed flow cytometrically for the determination of percent early apoptosis ( right panel ). (D) Cytotoxic effect of different doses of capsaisin on HUVEC cells were measured by trypan blue dye-exclusion assay. (E) Graphical representation of HUVEC migration upon incubation with spent media from capsaicin-treated (37.5 µM) HBL-100, HCT-15, HeLa and A549. (F) Representative images of capillary-like sprout formation by HUVECs in presence of media alone or spent media of WI-38/A549/Hy-A549/capsaicin-treated Hy-A549. Values are mean ± SEM of three independent experiments in each case or representative of typical experiment.

    Article Snippet: Cell culture Human cancer cell lines, A549, MCF-7, HeLa, HCT-15, and normal lung fibroblast WI-38, were obtained from National Centre for Cell Science, India, and maintained at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS (Lonza, NH), L-glutamine (2 mM), sodium pyruvate (100 µg/ml), nonessential amino acids (100 µM), streptomycin (100 µg/ml), penicillin (50 U/ml; Invitogen, CA) .

    Techniques: Migration, Wound Healing Assay, Incubation, Exclusion Assay, Binding Assay, Flow Cytometry

    Capsaicin inhibits VEGF transcriptional activation by targeting HIF-1α in a p53-dependent manner. (A) Time-dependent expression profiles of HIF-1α mRNA and -protein were determined by Western blot and RT-PCR, respectively, in capsaicin-treated Hy-A549 cells ( left panel ). Capsaicin-treated Hy-A549 cells were examined for time-dependent variation in the expression profiles of HIF-1α by quantitative real time PCR analysis and represented graphically ( right panel ). (B) Hy-A549 cells, transiently transfected with a non-targeting control-siRNA or HIF-1α-siRNA, were incubated with/without capsaicin for 24 h; the cells were then analyzed to determine VEGF expression at protein and mRNA levels ( left panel ). Immunoblot showing transfection efficiency of HIF-1α ( right panel ) (C) Control-siRNA/HIF-1α-siRNA-transfected Hy-A549 cell-supernatants were used to assess HUVEC migration by wound healing assay after capsaicin-treatment (37.5 µM; 24 h) and represented graphically. (D) Time-dependent expression profile of p53-mRNA and -protein was determined by Western blotting and RT-PCR in capsaicin-treated Hy-A549 cells ( left panel ). p53 phosphorylation at Serine-15 position was also evaluated ( right panel ). (E) Hy-A549 cells, transfected with control-shRNA/p53-shRNA were incubated with capsaicin for 24 h and HIF-1α VEGF-mRNA and -protein were determined by Western blot and RT-PCR ( left panel ). Transfection efficiency was checked by analyzing p53 expression level ( right panel ). (F) HIF-1α was immunoprecipitated from capsaicin-treated Hy-A549 cell lysates and immunoblotted with anti-Ub antibody to assay HIF-1α ubiquitination. The ladder of bands represented ubiquitinated HIF-1α. In parallel experiment, immunoprecipitates were assayed for HIF-1α levels by Western blot. Comparable protein input was confirmed by direct Western blotting with anti-α-actin using 20% of the cell lysates that were used for immunoprecipitation. (G) Control and MG-132 drug-pretreated Hy-A549 cells were subjected to capsaicin-treatment for 24 h and then were examined for expression of HIF-1α/VEGF by Western blotting. α-Actin/GAPDH was used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Journal: PLoS ONE

    Article Title: Capsaicin-Induced Activation of p53-SMAR1 Auto-Regulatory Loop Down-Regulates VEGF in Non-Small Cell Lung Cancer to Restrain Angiogenesis

    doi: 10.1371/journal.pone.0099743

    Figure Lengend Snippet: Capsaicin inhibits VEGF transcriptional activation by targeting HIF-1α in a p53-dependent manner. (A) Time-dependent expression profiles of HIF-1α mRNA and -protein were determined by Western blot and RT-PCR, respectively, in capsaicin-treated Hy-A549 cells ( left panel ). Capsaicin-treated Hy-A549 cells were examined for time-dependent variation in the expression profiles of HIF-1α by quantitative real time PCR analysis and represented graphically ( right panel ). (B) Hy-A549 cells, transiently transfected with a non-targeting control-siRNA or HIF-1α-siRNA, were incubated with/without capsaicin for 24 h; the cells were then analyzed to determine VEGF expression at protein and mRNA levels ( left panel ). Immunoblot showing transfection efficiency of HIF-1α ( right panel ) (C) Control-siRNA/HIF-1α-siRNA-transfected Hy-A549 cell-supernatants were used to assess HUVEC migration by wound healing assay after capsaicin-treatment (37.5 µM; 24 h) and represented graphically. (D) Time-dependent expression profile of p53-mRNA and -protein was determined by Western blotting and RT-PCR in capsaicin-treated Hy-A549 cells ( left panel ). p53 phosphorylation at Serine-15 position was also evaluated ( right panel ). (E) Hy-A549 cells, transfected with control-shRNA/p53-shRNA were incubated with capsaicin for 24 h and HIF-1α VEGF-mRNA and -protein were determined by Western blot and RT-PCR ( left panel ). Transfection efficiency was checked by analyzing p53 expression level ( right panel ). (F) HIF-1α was immunoprecipitated from capsaicin-treated Hy-A549 cell lysates and immunoblotted with anti-Ub antibody to assay HIF-1α ubiquitination. The ladder of bands represented ubiquitinated HIF-1α. In parallel experiment, immunoprecipitates were assayed for HIF-1α levels by Western blot. Comparable protein input was confirmed by direct Western blotting with anti-α-actin using 20% of the cell lysates that were used for immunoprecipitation. (G) Control and MG-132 drug-pretreated Hy-A549 cells were subjected to capsaicin-treatment for 24 h and then were examined for expression of HIF-1α/VEGF by Western blotting. α-Actin/GAPDH was used as internal loading control. Values are mean ±SEM of three independent experiments in each case or representative of typical experiment.

    Article Snippet: Cell culture Human cancer cell lines, A549, MCF-7, HeLa, HCT-15, and normal lung fibroblast WI-38, were obtained from National Centre for Cell Science, India, and maintained at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS (Lonza, NH), L-glutamine (2 mM), sodium pyruvate (100 µg/ml), nonessential amino acids (100 µM), streptomycin (100 µg/ml), penicillin (50 U/ml; Invitogen, CA) .

    Techniques: Activation Assay, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Transfection, Incubation, Migration, Wound Healing Assay, shRNA, Immunoprecipitation

    Capsaicin inhibits the nuclear localization of HIF-1α by down regulating Cox-2 in a p53-dependent manner. (A) Immunoblot representing nuclear and cytosolic levels of HIF-1α in capsaicin-treated/p53-shRNA-transfected Hy-A549 cells. (B) HIF-1α expression was monitored in capsaicin-treated Hy-A549 cells by confocal microscopy (magnification 60x). (C) Time-dependent expression of Cox-2-mRNA and -protein in capsaicin-treated Hy-A549 cells was determined by Western blot and RT-PCR. (D) Control siRNA-/Cox-2-siRNA-transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and nuclear translocation of HIF-1α was assessed by Western blot analysis ( left panel ). Transfection efficiency was determined by analyzing the expression of Cox-2 ( right panel ). (E) Control vector-/Cox-2 cDNA transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and analyzed for nuclear and cytosolic expression of HIF-1α ( left panel ). Transfection efficiency of Cox-2 was also verified ( right panel ). (F) Control-shRNA-/p53-shRNA-transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and expression profiles of Cox-2 both at protein and mRNA level were examined. (G) Time-dependent expression profiles of SMAR1-protein and mRNA in capsaicin-treated Hy-A549 cells were determined by Western blot and RT-PCR ( left panels ). Control-/p53-shRNA transfected Hy-A549 cells were treated with capsaicin and expression levels of SMAR1 were checked ( right panel ). (H) Control-/SMAR1-shRNA transfected Hy-A549 cells were treated with capsaicin and immune-blotted with p-Ser 15 -p53. α-Actin/H1-Histone/GAPDH were used as internal loading control. Values are mean ± SEM of three independent experiments in each case or representative of typical experiment.

    Journal: PLoS ONE

    Article Title: Capsaicin-Induced Activation of p53-SMAR1 Auto-Regulatory Loop Down-Regulates VEGF in Non-Small Cell Lung Cancer to Restrain Angiogenesis

    doi: 10.1371/journal.pone.0099743

    Figure Lengend Snippet: Capsaicin inhibits the nuclear localization of HIF-1α by down regulating Cox-2 in a p53-dependent manner. (A) Immunoblot representing nuclear and cytosolic levels of HIF-1α in capsaicin-treated/p53-shRNA-transfected Hy-A549 cells. (B) HIF-1α expression was monitored in capsaicin-treated Hy-A549 cells by confocal microscopy (magnification 60x). (C) Time-dependent expression of Cox-2-mRNA and -protein in capsaicin-treated Hy-A549 cells was determined by Western blot and RT-PCR. (D) Control siRNA-/Cox-2-siRNA-transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and nuclear translocation of HIF-1α was assessed by Western blot analysis ( left panel ). Transfection efficiency was determined by analyzing the expression of Cox-2 ( right panel ). (E) Control vector-/Cox-2 cDNA transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and analyzed for nuclear and cytosolic expression of HIF-1α ( left panel ). Transfection efficiency of Cox-2 was also verified ( right panel ). (F) Control-shRNA-/p53-shRNA-transfected Hy-A549 cells were treated with capsaicin (37.5 µM; 24 h) and expression profiles of Cox-2 both at protein and mRNA level were examined. (G) Time-dependent expression profiles of SMAR1-protein and mRNA in capsaicin-treated Hy-A549 cells were determined by Western blot and RT-PCR ( left panels ). Control-/p53-shRNA transfected Hy-A549 cells were treated with capsaicin and expression levels of SMAR1 were checked ( right panel ). (H) Control-/SMAR1-shRNA transfected Hy-A549 cells were treated with capsaicin and immune-blotted with p-Ser 15 -p53. α-Actin/H1-Histone/GAPDH were used as internal loading control. Values are mean ± SEM of three independent experiments in each case or representative of typical experiment.

    Article Snippet: Cell culture Human cancer cell lines, A549, MCF-7, HeLa, HCT-15, and normal lung fibroblast WI-38, were obtained from National Centre for Cell Science, India, and maintained at 37°C and 5% CO2 in DMEM medium supplemented with 10% FBS (Lonza, NH), L-glutamine (2 mM), sodium pyruvate (100 µg/ml), nonessential amino acids (100 µM), streptomycin (100 µg/ml), penicillin (50 U/ml; Invitogen, CA) .

    Techniques: shRNA, Transfection, Expressing, Confocal Microscopy, Western Blot, Reverse Transcription Polymerase Chain Reaction, Translocation Assay, Plasmid Preparation

    MTT analysis with different concentration of Caesalpinia sappan L. leaf and heart wood extracts in chloroform, water and methanol on A549 (Human lung cancer) cell line. *L – leaf and H – heartwood. .

    Journal: Data in Brief

    Article Title: In vitro studies data on anticancer activity of Caesalpinia sappan L. heartwood and leaf extracts on MCF7 and A549 cell lines

    doi: 10.1016/j.dib.2018.05.050

    Figure Lengend Snippet: MTT analysis with different concentration of Caesalpinia sappan L. leaf and heart wood extracts in chloroform, water and methanol on A549 (Human lung cancer) cell line. *L – leaf and H – heartwood. .

    Article Snippet: The pure cultures of MCF-7 (Breast cancer cell line) and A549 ( Lung Cancer), were obtained From National Centre for Cell Science, Pune, Maharashtra state,India.

    Techniques: MTT Assay, Concentration Assay

    Nimbolide interacts with TNF-α and inhibits TNF-α regulated inflammatory signaling. a Docking model of nimbolide in the active site of TNF-α (PDB ID: 2AZ5) and ( b ) its ligand-protein interactions in the binding site of TNF-α. The dark pink dashed lines represent hydrogen bonds. H-bond distances (in Å) between heteroatoms of ligand and amino acid residues are as follows: Ser60 (3.4Å) and Leu120 (3.2Å). The red line indicates arene-arene interaction with Tyr59. A549 cells were pre-treated with nimbolide for 24 h and stimulated by LPS (1 µg/ml) for 12 h. Animals were pre-treated with nimbolide (0.3, 1 and 3 mg/kg) for 5 days later LPS (50 μg) was administered. c TNF-α protein expression was analyzed by confocal microscope. d A 5-µm-sized sections of lung tissues were subjected to IHC to determine TNF-α expression and images were captured at ×400 magnification. Protein expressions of TNF-α, p-p38 MAPK, p-IKK-α/β, p-IκB-α, p-NF-κB, p-GSK-3β, and mTOR were determined by western blotting in ( e ) A549 cells and ( f ) lung tissues, respectively. The HDACs such as HDAC-1, 2, 3, and 4 protein expressions were studied in ( g ) A549 cells and ( h ) lung tissues. i Nimbolide (0.05, 0.1, 1, and 2.5 µM) upon HDAC levels were analyzed by HDAC fluorometric kit and HDAC inhibitory activity was compared with trichostatin A (2.5 µM). Data represented as mean ± SEM (n=3 independent experiments). * P

    Journal: Cell Death & Disease

    Article Title: Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation

    doi: 10.1038/s41419-018-1247-9

    Figure Lengend Snippet: Nimbolide interacts with TNF-α and inhibits TNF-α regulated inflammatory signaling. a Docking model of nimbolide in the active site of TNF-α (PDB ID: 2AZ5) and ( b ) its ligand-protein interactions in the binding site of TNF-α. The dark pink dashed lines represent hydrogen bonds. H-bond distances (in Å) between heteroatoms of ligand and amino acid residues are as follows: Ser60 (3.4Å) and Leu120 (3.2Å). The red line indicates arene-arene interaction with Tyr59. A549 cells were pre-treated with nimbolide for 24 h and stimulated by LPS (1 µg/ml) for 12 h. Animals were pre-treated with nimbolide (0.3, 1 and 3 mg/kg) for 5 days later LPS (50 μg) was administered. c TNF-α protein expression was analyzed by confocal microscope. d A 5-µm-sized sections of lung tissues were subjected to IHC to determine TNF-α expression and images were captured at ×400 magnification. Protein expressions of TNF-α, p-p38 MAPK, p-IKK-α/β, p-IκB-α, p-NF-κB, p-GSK-3β, and mTOR were determined by western blotting in ( e ) A549 cells and ( f ) lung tissues, respectively. The HDACs such as HDAC-1, 2, 3, and 4 protein expressions were studied in ( g ) A549 cells and ( h ) lung tissues. i Nimbolide (0.05, 0.1, 1, and 2.5 µM) upon HDAC levels were analyzed by HDAC fluorometric kit and HDAC inhibitory activity was compared with trichostatin A (2.5 µM). Data represented as mean ± SEM (n=3 independent experiments). * P

    Article Snippet: Cell culture RAW 264.7 (murine macrophages) and A549 (human type II alveolar epithelial cells) cells were obtained from National Centre for Cell Science (NCCS), Pune, India.

    Techniques: Binding Assay, Expressing, Microscopy, Immunohistochemistry, Western Blot, Activity Assay

    Nimbolide suppresses NF-κB and HDAC-3 protein expression and nuclear translocation. A549 cells were pre-treated with nimbolide for 24 h. After 30 min of LPS (1 µg/ml) incubation, both cytosolic and nuclear proteins were isolated. NF-κB and HDAC-3 protein levels were observed in ( a ) cytosolic and ( b ) nuclear fraction of A549 cells. c Similarly, the protein expression of NF-κB and HDAC-3 was analyzed by confocal microscope in A549 cells. d Microsections of lung tissues were subjected to IHC to determine NF-κB and HDAC-3 expressions and images were captured at ×400 magnification

    Journal: Cell Death & Disease

    Article Title: Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation

    doi: 10.1038/s41419-018-1247-9

    Figure Lengend Snippet: Nimbolide suppresses NF-κB and HDAC-3 protein expression and nuclear translocation. A549 cells were pre-treated with nimbolide for 24 h. After 30 min of LPS (1 µg/ml) incubation, both cytosolic and nuclear proteins were isolated. NF-κB and HDAC-3 protein levels were observed in ( a ) cytosolic and ( b ) nuclear fraction of A549 cells. c Similarly, the protein expression of NF-κB and HDAC-3 was analyzed by confocal microscope in A549 cells. d Microsections of lung tissues were subjected to IHC to determine NF-κB and HDAC-3 expressions and images were captured at ×400 magnification

    Article Snippet: Cell culture RAW 264.7 (murine macrophages) and A549 (human type II alveolar epithelial cells) cells were obtained from National Centre for Cell Science (NCCS), Pune, India.

    Techniques: Expressing, Translocation Assay, Incubation, Isolation, Microscopy, Immunohistochemistry

    Nimbolide inhibits TNF-α regulated NF-κB and HDAC-3 protein expression and nuclear translocation. A549 cells were treated with nimbolide (2.5 µM) for 24 h and stimulated with TNF-α (10 ng/ml) for 30 min. a NF-κB and HDAC-3 protein levels were observed in both cytosolic and nuclear fraction of A549 cells by western blotting. b The confocal analysis was performed to determine the protein expressions of NF-κB and HDAC-3. All the images were captured at ×400 magnification. BEAS-2B cells were transfected with TNF-α and scrambled siRNA and incubated for 24 h. In another set of the experimental group, cells were pre-treated with nimbolide (2.5 µM) for 24 h. Then, cells were stimulated with TNF-α (10 ng/ml) for 30 min except for nimbolide alone (NIM) group (2.5 µM). The expression of TNF-α, NF-κB and HDAC-3 levels were measured by ( c ) western blotting and ( d ) confocal analysis. The images were captured at ×400 magnification

    Journal: Cell Death & Disease

    Article Title: Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation

    doi: 10.1038/s41419-018-1247-9

    Figure Lengend Snippet: Nimbolide inhibits TNF-α regulated NF-κB and HDAC-3 protein expression and nuclear translocation. A549 cells were treated with nimbolide (2.5 µM) for 24 h and stimulated with TNF-α (10 ng/ml) for 30 min. a NF-κB and HDAC-3 protein levels were observed in both cytosolic and nuclear fraction of A549 cells by western blotting. b The confocal analysis was performed to determine the protein expressions of NF-κB and HDAC-3. All the images were captured at ×400 magnification. BEAS-2B cells were transfected with TNF-α and scrambled siRNA and incubated for 24 h. In another set of the experimental group, cells were pre-treated with nimbolide (2.5 µM) for 24 h. Then, cells were stimulated with TNF-α (10 ng/ml) for 30 min except for nimbolide alone (NIM) group (2.5 µM). The expression of TNF-α, NF-κB and HDAC-3 levels were measured by ( c ) western blotting and ( d ) confocal analysis. The images were captured at ×400 magnification

    Article Snippet: Cell culture RAW 264.7 (murine macrophages) and A549 (human type II alveolar epithelial cells) cells were obtained from National Centre for Cell Science (NCCS), Pune, India.

    Techniques: Expressing, Translocation Assay, Western Blot, Transfection, Incubation