hep 2 cells  (ATCC)


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    Name:
    HEp 2
    Description:

    Catalog Number:
    ccl-23
    Price:
    None
    Host:
    Homo sapiens, human
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    Structured Review

    ATCC hep 2 cells
    Cellular distribution and orientation of GFP-RSV F tail in nonpolarized epithelial cells. (a) <t>HEp-2</t> cells were infected with RSV for 48 h to determine the cellular distribution of the F protein. RSV F (red) was distributed in a perinuclear manner, consistent

    https://www.bioz.com/result/hep 2 cells/product/ATCC
    Average 99 stars, based on 43 article reviews
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    Images

    1) Product Images from "The Transmembrane Domain of the Respiratory Syncytial Virus F Protein Is an Orientation-Independent Apical Plasma Membrane Sorting Sequence"

    Article Title: The Transmembrane Domain of the Respiratory Syncytial Virus F Protein Is an Orientation-Independent Apical Plasma Membrane Sorting Sequence

    Journal:

    doi: 10.1128/JVI.79.19.12528-12535.2005

    Cellular distribution and orientation of GFP-RSV F tail in nonpolarized epithelial cells. (a) HEp-2 cells were infected with RSV for 48 h to determine the cellular distribution of the F protein. RSV F (red) was distributed in a perinuclear manner, consistent
    Figure Legend Snippet: Cellular distribution and orientation of GFP-RSV F tail in nonpolarized epithelial cells. (a) HEp-2 cells were infected with RSV for 48 h to determine the cellular distribution of the F protein. RSV F (red) was distributed in a perinuclear manner, consistent

    Techniques Used: Infection

    ER and Golgi colocalization of GFP-RSV F tail constructs in nonpolarized HEp-2 cells. HEp-2 cells were transfected with GFP-RSV F tail (a and b) or GFP-RSV F tail (ΔCT) (c and d) to assess the codistribution of each protein with resident ER (a and
    Figure Legend Snippet: ER and Golgi colocalization of GFP-RSV F tail constructs in nonpolarized HEp-2 cells. HEp-2 cells were transfected with GFP-RSV F tail (a and b) or GFP-RSV F tail (ΔCT) (c and d) to assess the codistribution of each protein with resident ER (a and

    Techniques Used: Construct, Transfection

    Cellular distribution of RSV F opt in nonpolarized epithelial cells. (a) Schematic representation of full-length and mutant forms of RSV F opt . (b to e) HEp-2 cells were transfected with each RSV F opt construct noted in panel a to assess the cellular distribution
    Figure Legend Snippet: Cellular distribution of RSV F opt in nonpolarized epithelial cells. (a) Schematic representation of full-length and mutant forms of RSV F opt . (b to e) HEp-2 cells were transfected with each RSV F opt construct noted in panel a to assess the cellular distribution

    Techniques Used: Mutagenesis, Transfection, Construct

    2) Product Images from "The Carboxyl-Terminal Domain of RNA Polymerase II Is Phosphorylated by a Complex Containing cdk9 and Infected-Cell Protein 22 of Herpes Simplex Virus 1"

    Article Title: The Carboxyl-Terminal Domain of RNA Polymerase II Is Phosphorylated by a Complex Containing cdk9 and Infected-Cell Protein 22 of Herpes Simplex Virus 1

    Journal:

    doi: 10.1128/JVI.79.11.6757-6762.2005

    ICP22-dependent phosphorylation of RNA POL II CTD requires the US3 protein kinase. (A and D) HEp-2 cells were mock infected or exposed to HSV-1(F), R325 (Δα22), R7356 (ΔU L 13), R7041 (ΔU S 3), or R7353 (ΔU L 13/ΔU
    Figure Legend Snippet: ICP22-dependent phosphorylation of RNA POL II CTD requires the US3 protein kinase. (A and D) HEp-2 cells were mock infected or exposed to HSV-1(F), R325 (Δα22), R7356 (ΔU L 13), R7041 (ΔU S 3), or R7353 (ΔU L 13/ΔU

    Techniques Used: Infection

    GST-cdk9 interacts with ICP22. Lysates of HEp-2 cells mock infected or infected with HSV-1(F) or R325 (ΔICP22) and incubated for 6 h were reacted with GST-cdk9. The bound proteins were solubilized, subjected to electrophoresis on a denaturing
    Figure Legend Snippet: GST-cdk9 interacts with ICP22. Lysates of HEp-2 cells mock infected or infected with HSV-1(F) or R325 (ΔICP22) and incubated for 6 h were reacted with GST-cdk9. The bound proteins were solubilized, subjected to electrophoresis on a denaturing

    Techniques Used: Infection, Incubation, Electrophoresis

    The interaction of GST-cdk9 chimeric protein with the 69,000- M r protein is dependent on ICP22 and the U S 3 protein kinase. Panel A: Autoradiogram of electrophoretically separated lysates of [ 35 S]Met-labeled HEp-2 cell proteins. The cells were harvested
    Figure Legend Snippet: The interaction of GST-cdk9 chimeric protein with the 69,000- M r protein is dependent on ICP22 and the U S 3 protein kinase. Panel A: Autoradiogram of electrophoretically separated lysates of [ 35 S]Met-labeled HEp-2 cell proteins. The cells were harvested

    Techniques Used: Labeling

    3) Product Images from "RNA-associated protein 55 (RAP55) localizes to mRNA processing bodies and stress granules"

    Article Title: RNA-associated protein 55 (RAP55) localizes to mRNA processing bodies and stress granules

    Journal:

    doi: 10.1261/rna.2302706

    Antibodies in the serum of a primary biliary cirrhosis patient react with P-bodies and stress granules. (Panel I ) ( A,E ) Patient 0080’s serum contained antibodies that reacted with 5–20 dots in the cytoplasm of Hep-2 cells as determined
    Figure Legend Snippet: Antibodies in the serum of a primary biliary cirrhosis patient react with P-bodies and stress granules. (Panel I ) ( A,E ) Patient 0080’s serum contained antibodies that reacted with 5–20 dots in the cytoplasm of Hep-2 cells as determined

    Techniques Used:

    The effect of arsenite-induced oxidative stress on the cellular location of RAP55. ( A ) In resting Hep-2 cells, RAP55-containing P-bodies were distributed throughout the cytoplasm, and ( B ) TIA was detected diffusely throughout the nucleus, but excluded
    Figure Legend Snippet: The effect of arsenite-induced oxidative stress on the cellular location of RAP55. ( A ) In resting Hep-2 cells, RAP55-containing P-bodies were distributed throughout the cytoplasm, and ( B ) TIA was detected diffusely throughout the nucleus, but excluded

    Techniques Used:

    The effect of RAP55 knock-down on P-bodies. (Panel I ) Immunoblotting was used to determine the effect of siRNAs on the levels of RAP55 and DCP1a in Hep-2 cells. Compared with control siRNA, RAP55 siRNA decreased the level of ( A ) RAP55 but not ( B ) DCP1a
    Figure Legend Snippet: The effect of RAP55 knock-down on P-bodies. (Panel I ) Immunoblotting was used to determine the effect of siRNAs on the levels of RAP55 and DCP1a in Hep-2 cells. Compared with control siRNA, RAP55 siRNA decreased the level of ( A ) RAP55 but not ( B ) DCP1a

    Techniques Used:

    4) Product Images from "Translocation and Colocalization of ICP4 and ICP0 in Cells Infected with Herpes Simplex Virus 1 Mutants Lacking Glycoprotein E, Glycoprotein I, or the Virion Host Shutoff Product of the UL41 Gene ▿"

    Article Title: Translocation and Colocalization of ICP4 and ICP0 in Cells Infected with Herpes Simplex Virus 1 Mutants Lacking Glycoprotein E, Glycoprotein I, or the Virion Host Shutoff Product of the UL41 Gene ▿

    Journal:

    doi: 10.1128/JVI.02157-07

    Properties of the ΔgE, ΔgI, and ΔU L 41 viruses. (A and B) Expression patterns of gE and gI in ΔU L 41 virus-infected cells. HEp-2 cells were either mock infected or exposed (10 PFU/cell) to HSV-1(F), ΔU L 41, ΔgE-2
    Figure Legend Snippet: Properties of the ΔgE, ΔgI, and ΔU L 41 viruses. (A and B) Expression patterns of gE and gI in ΔU L 41 virus-infected cells. HEp-2 cells were either mock infected or exposed (10 PFU/cell) to HSV-1(F), ΔU L 41, ΔgE-2

    Techniques Used: Expressing, Infection

    Exportation of ICP4 from the nucleus and aggregation in the cytoplasm of the ΔgE and ΔU L 41 virus-infected cells. Hep-2 cells, seeded in four-well slides, were either mock infected or exposed (10 PFU/cell) to HSV-1(F), ΔgE, or ΔU
    Figure Legend Snippet: Exportation of ICP4 from the nucleus and aggregation in the cytoplasm of the ΔgE and ΔU L 41 virus-infected cells. Hep-2 cells, seeded in four-well slides, were either mock infected or exposed (10 PFU/cell) to HSV-1(F), ΔgE, or ΔU

    Techniques Used: Infection

    Electron photomicrographs of ICP0-containing structures in HSV-1(F)- and ΔgE-infected cells. HEp-2 cells were harvested at 18 h after infection with 10 PFU/cell of HSV-1(F) or the ΔgE mutant and processed for immunoelectron microscopy,
    Figure Legend Snippet: Electron photomicrographs of ICP0-containing structures in HSV-1(F)- and ΔgE-infected cells. HEp-2 cells were harvested at 18 h after infection with 10 PFU/cell of HSV-1(F) or the ΔgE mutant and processed for immunoelectron microscopy,

    Techniques Used: Infection, Mutagenesis, Immuno-Electron Microscopy

    5) Product Images from "O-Linked Glycosylation Ensures the Normal Conformation of the Autotransporter Adhesin Involved in Diffuse Adherence ▿"

    Article Title: O-Linked Glycosylation Ensures the Normal Conformation of the Autotransporter Adhesin Involved in Diffuse Adherence ▿

    Journal:

    doi: 10.1128/JB.00969-07

    Effect of glycosylation on the function of AIDA-I. (A) Adhesion assay. Bacteria bearing an empty vector (−) or expressing unglycosylated AIDA-I (Aida) or glycosylated AIDA-I (Aida/Aah) were inoculated onto a monolayer of confluent Hep-2 cells,
    Figure Legend Snippet: Effect of glycosylation on the function of AIDA-I. (A) Adhesion assay. Bacteria bearing an empty vector (−) or expressing unglycosylated AIDA-I (Aida) or glycosylated AIDA-I (Aida/Aah) were inoculated onto a monolayer of confluent Hep-2 cells,

    Techniques Used: Cell Adhesion Assay, Plasmid Preparation, Expressing

    Related Articles

    Transformation Assay:

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Disrupts Stat1-Mediated Gamma Interferon Signal Transduction in Epithelial Cells
    Article Snippet: .. The transformed human laryngeal epithelial cell line HEp-2 (= ATCC CCL-23) was cultured in minimum essential medium supplemented with 15% fetal bovine serum (FBS), 2.5% penicillin-streptomycin, 1.8% sodium bicarbonate, and 1.2% Fungizone (all obtained from Life Technologies, Grand Island, N.Y.) ( ). .. The transformed human colonic epithelial cell line T84 (= ATCC CCL-248; American Type Culture Collection, Manassas, Va.) was cultured in a 1:1 mixture of Dulbecco's modified Eagle medium and Ham's F-12 medium supplemented with 10% (vol/vol) FBS, 2% penicillin-streptomycin, 2% sodium bicarbonate, and 0.6% l -glutamine ( ).

    other:

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Shiga Toxins Inhibit Gamma Interferon-Mediated Cellular Activation
    Article Snippet: HEp-2 epithelial cells (ATCC CCL-23) were used as a model epithelial cell line, as previously described ( ).

    Article Title: Herpes simplex virus protein kinase US3 activates and functionally overlaps protein kinase A to block apoptosis
    Article Snippet: SK-N-SH and HEp-2 cell lines obtained from the American Type Culture Collection were grown in DMEM containing 5% (HEp-2) or 10% (SK-N-SH) newborn calf serum.

    Article Title: Chlamydia pneumoniae Promotes Dysfunction of Pancreatic Beta Cells
    Article Snippet: However, mast cells and Hep2 cells were susceptible at 1 MOI.

    Cell Culture:

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Disrupts Stat1-Mediated Gamma Interferon Signal Transduction in Epithelial Cells
    Article Snippet: .. The transformed human laryngeal epithelial cell line HEp-2 (= ATCC CCL-23) was cultured in minimum essential medium supplemented with 15% fetal bovine serum (FBS), 2.5% penicillin-streptomycin, 1.8% sodium bicarbonate, and 1.2% Fungizone (all obtained from Life Technologies, Grand Island, N.Y.) ( ). .. The transformed human colonic epithelial cell line T84 (= ATCC CCL-248; American Type Culture Collection, Manassas, Va.) was cultured in a 1:1 mixture of Dulbecco's modified Eagle medium and Ham's F-12 medium supplemented with 10% (vol/vol) FBS, 2% penicillin-streptomycin, 2% sodium bicarbonate, and 0.6% l -glutamine ( ).

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  • 99
    ATCC hk 2 cells human kidney cortex proximal tubule epithelial cells
    Cell protection capabilities of rA1M. (A) K562 cells, seeded at 10 5 cells per well in a 96-well microtiter plate, were exposed to 100 μ M heme in the presence of a dilution series (0–10 μ M ) of rA1M-wt- (▪), rA1M-035 (○), or ovalbumin (●) for 1 h. Cell death was monitored as release of LDH into the medium. The LDH value from live cells was subtracted and the signal of heme-incubated cells without rA1M was set to 100% and the values of the rA1M incubations were calculated in relation to this. The assay was made in duplicate. The average result from three independent experiments (mean ± SEM) is shown. (B–E) <t>HK-2</t> cells were exposed to a mixture of 200 μ M (NH 4 )Fe(SO 4 ) 2 , 400 μ M hydrogen peroxide, and 2 m M ascorbate (the Fenton reaction, B and C ), 10 or 30 μ M heme ( D ), and 30 μ M heme (E) with or without the simultaneous addition of 0–20 μ M rA1M-wt (displayed as ▪ in B and D , and black columns in C and E ) or rA1M-035 (displayed as ○ in B and D , and white columns in C and E ) for 6 h. After incubation, cells were analyzed for cell viability using (B and D) WST-1 or (C, E) mRNA expression of HO-1 and Hsp70. The cell viability ( B and D ) was normalized against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. The mRNA expression of HO-1 and Hsp70 (C, E) was normalized against GAPDH and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures and control conditions were analyzed using one-way ANOVA with post hoc Bonferroni correction. *Statistical comparison versus (C) Fenton or (E) heme. *** p
    Hk 2 Cells Human Kidney Cortex Proximal Tubule Epithelial Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hk 2 cells human kidney cortex proximal tubule epithelial cells/product/ATCC
    Average 99 stars, based on 1 article reviews
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    89
    ATCC sk hep 1
    p62 KD <t>SK-HEP-1</t> and p62 KD MDA-MB-231 cells exhibit increased cell migration in response to TLR4 stimulation. ( A , B ) Ctrl and p62 KD SK-HEP-1 cells were seeded into 12-well cell culture plates. Confluent monolayers were scraped with a sterile yellow Gilson-pipette tip, and the wound was then treated with vehicle (DMSO,
    Sk Hep 1, supplied by ATCC, used in various techniques. Bioz Stars score: 89/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    ATCC transfectionthe human pancreatic epithelial cell line mia paca 2
    c-MYC associated with PCAF and CtBPs to assemble a complex. (A) The Flag-c-MYC-associated complex. The pCDNA3-2×Flag (empty vector, EV) and pCDNA3-2×Flag-c-MYC plasmids were transfected into MIA <t>PaCa-2</t> cells, respectively. The resulting cells were subjected to immunoprecipitation with the anti-Flag resin. The purified complexes were separated in an SDS-PAGE gel and incubated with a silver staining kit. The IgG and Flag-c-MYC were indicated by arrows. (B) c-MYC could pull down PCAF and CtBPs in vivo . Equal weight of pancreatic tissues from three AP patients was mixed and lysed, and 1/11 cell extracts were used as an input, and the other 10/11 cell extracts were equally divided into two parts, followed by immunoprecipitation with an IgG and anti-c-MYC antibody-associated protein A beads, respectively. The input and output proteins were used to determine protein levels of c-MYC, PCAF, CtBP1 and CtBP2, respectively. (C) c-MYC directly interacted with PCAF but not CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-PCAF, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-PCAF. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies. (D) PCAF directly interacted with both c-MYC and CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output protein levels using anti-Flag and anti-Myc antibodies. (E) CtBPs assembled a heterotetramer in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies.
    Transfectionthe Human Pancreatic Epithelial Cell Line Mia Paca 2, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    ATCC confluent hep 2 cells
    Continuous C. pneumoniae infection in <t>HEp-2</t> cells inoculated with isolate CM-1 (A to D) or isolate TW-183 (E and F). (A and B) Typical inclusions; (C to F) altered inclusions; (D) aberrant inclusion. C, cytoplasm; N, nucleus; M, mitochondria; im, inclusion membrane; e, intrainclusional membranous material.
    Confluent Hep 2 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell protection capabilities of rA1M. (A) K562 cells, seeded at 10 5 cells per well in a 96-well microtiter plate, were exposed to 100 μ M heme in the presence of a dilution series (0–10 μ M ) of rA1M-wt- (▪), rA1M-035 (○), or ovalbumin (●) for 1 h. Cell death was monitored as release of LDH into the medium. The LDH value from live cells was subtracted and the signal of heme-incubated cells without rA1M was set to 100% and the values of the rA1M incubations were calculated in relation to this. The assay was made in duplicate. The average result from three independent experiments (mean ± SEM) is shown. (B–E) HK-2 cells were exposed to a mixture of 200 μ M (NH 4 )Fe(SO 4 ) 2 , 400 μ M hydrogen peroxide, and 2 m M ascorbate (the Fenton reaction, B and C ), 10 or 30 μ M heme ( D ), and 30 μ M heme (E) with or without the simultaneous addition of 0–20 μ M rA1M-wt (displayed as ▪ in B and D , and black columns in C and E ) or rA1M-035 (displayed as ○ in B and D , and white columns in C and E ) for 6 h. After incubation, cells were analyzed for cell viability using (B and D) WST-1 or (C, E) mRNA expression of HO-1 and Hsp70. The cell viability ( B and D ) was normalized against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. The mRNA expression of HO-1 and Hsp70 (C, E) was normalized against GAPDH and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures and control conditions were analyzed using one-way ANOVA with post hoc Bonferroni correction. *Statistical comparison versus (C) Fenton or (E) heme. *** p

    Journal: Antioxidants & Redox Signaling

    Article Title: rA1M-035, a Physicochemically Improved Human Recombinant α1-Microglobulin, Has Therapeutic Effects in Rhabdomyolysis-Induced Acute Kidney Injury

    doi: 10.1089/ars.2017.7181

    Figure Lengend Snippet: Cell protection capabilities of rA1M. (A) K562 cells, seeded at 10 5 cells per well in a 96-well microtiter plate, were exposed to 100 μ M heme in the presence of a dilution series (0–10 μ M ) of rA1M-wt- (▪), rA1M-035 (○), or ovalbumin (●) for 1 h. Cell death was monitored as release of LDH into the medium. The LDH value from live cells was subtracted and the signal of heme-incubated cells without rA1M was set to 100% and the values of the rA1M incubations were calculated in relation to this. The assay was made in duplicate. The average result from three independent experiments (mean ± SEM) is shown. (B–E) HK-2 cells were exposed to a mixture of 200 μ M (NH 4 )Fe(SO 4 ) 2 , 400 μ M hydrogen peroxide, and 2 m M ascorbate (the Fenton reaction, B and C ), 10 or 30 μ M heme ( D ), and 30 μ M heme (E) with or without the simultaneous addition of 0–20 μ M rA1M-wt (displayed as ▪ in B and D , and black columns in C and E ) or rA1M-035 (displayed as ○ in B and D , and white columns in C and E ) for 6 h. After incubation, cells were analyzed for cell viability using (B and D) WST-1 or (C, E) mRNA expression of HO-1 and Hsp70. The cell viability ( B and D ) was normalized against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. The mRNA expression of HO-1 and Hsp70 (C, E) was normalized against GAPDH and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures and control conditions were analyzed using one-way ANOVA with post hoc Bonferroni correction. *Statistical comparison versus (C) Fenton or (E) heme. *** p

    Article Snippet: Protection of HK-2 cells Human kidney cortex proximal tubule epithelial cells (HK-2, ATCC CRL-2190, ATCC, United Kingdom) were cultured in keratinocyte serum-free medium supplemented with bovine pituitary extract (0.05 mg/mL) and epidermal growth factor (5 ng/mL) (all from Invitrogen, United Kingdom).

    Techniques: Incubation, Expressing

    p62 KD SK-HEP-1 and p62 KD MDA-MB-231 cells exhibit increased cell migration in response to TLR4 stimulation. ( A , B ) Ctrl and p62 KD SK-HEP-1 cells were seeded into 12-well cell culture plates. Confluent monolayers were scraped with a sterile yellow Gilson-pipette tip, and the wound was then treated with vehicle (DMSO,

    Journal: Cells

    Article Title: p62 is Negatively Implicated in the TRAF6-BECN1 Signaling Axis for Autophagy Activation and Cancer Progression by Toll-Like Receptor 4 (TLR4)

    doi: 10.3390/cells9051142

    Figure Lengend Snippet: p62 KD SK-HEP-1 and p62 KD MDA-MB-231 cells exhibit increased cell migration in response to TLR4 stimulation. ( A , B ) Ctrl and p62 KD SK-HEP-1 cells were seeded into 12-well cell culture plates. Confluent monolayers were scraped with a sterile yellow Gilson-pipette tip, and the wound was then treated with vehicle (DMSO,

    Article Snippet: Human hepatic adenocarcinoma cell line SK-HEP-1 (ATCC, HTB-52), human breast adenocarcinoma cell line MDA-MB-231 (ATCC, HTB-26), and human lung cancer cell line A549 (ATCC, CCL-185) were purchased from ATCC, and cultured in DMEM or RPMI contained with 10% FBS.

    Techniques: Multiple Displacement Amplification, Migration, Cell Culture, Transferring

    p62 KD SK-HEP-1 and p62 KD MDA-MB-231 cells exhibit increased invasiveness in response to TLR4 stimulation. ( A , B ) Ctrl and p62 KD SK-HEP-1 cells were suspended in DMEM culture medium including vehicle, LPS (10 μg/mL), 3-MA (5 mM) plus LPS (10 μg/mL), and CQ (10 μM) plus LPS (10 μg/mL). Cells were placed into the top chambers of 24-transwell plates and incubated for overnight. Fixed cells were stained by using crystal violet ( A ). Numbers of migrated cells were counted, and results are represented as mean ± SEM ( B ). * p

    Journal: Cells

    Article Title: p62 is Negatively Implicated in the TRAF6-BECN1 Signaling Axis for Autophagy Activation and Cancer Progression by Toll-Like Receptor 4 (TLR4)

    doi: 10.3390/cells9051142

    Figure Lengend Snippet: p62 KD SK-HEP-1 and p62 KD MDA-MB-231 cells exhibit increased invasiveness in response to TLR4 stimulation. ( A , B ) Ctrl and p62 KD SK-HEP-1 cells were suspended in DMEM culture medium including vehicle, LPS (10 μg/mL), 3-MA (5 mM) plus LPS (10 μg/mL), and CQ (10 μM) plus LPS (10 μg/mL). Cells were placed into the top chambers of 24-transwell plates and incubated for overnight. Fixed cells were stained by using crystal violet ( A ). Numbers of migrated cells were counted, and results are represented as mean ± SEM ( B ). * p

    Article Snippet: Human hepatic adenocarcinoma cell line SK-HEP-1 (ATCC, HTB-52), human breast adenocarcinoma cell line MDA-MB-231 (ATCC, HTB-26), and human lung cancer cell line A549 (ATCC, CCL-185) were purchased from ATCC, and cultured in DMEM or RPMI contained with 10% FBS.

    Techniques: Multiple Displacement Amplification, Incubation, Staining

    p62-deficient cells, p62 KD THP-1, p62 KD SK-HEP-1, and p62 KD MDA-MB-231 cells, exhibit enhanced autophagy activation in response to TLR4 stimulation. ( A ) p62 KD THP-1 cells were generated, and the knockdown efficacy of p62 was confirmed with anti-p62 antibody. ( B , C ) Ctrl and p62 KD THP-1 cells were treated with or without vehicle or CQ (10 μM), in the presence or absence of LPS (10 μg/mL), for 6 h. Whole cell lysates were immunoblotted with anti-LC3A/B antibody and anti-GAPDH antibody as a loading control ( B ). The LC3II levels were analyzed with Image J program ( C ). Data shown are averages from a minimum of 3 independent experiments (± SEM). *, p

    Journal: Cells

    Article Title: p62 is Negatively Implicated in the TRAF6-BECN1 Signaling Axis for Autophagy Activation and Cancer Progression by Toll-Like Receptor 4 (TLR4)

    doi: 10.3390/cells9051142

    Figure Lengend Snippet: p62-deficient cells, p62 KD THP-1, p62 KD SK-HEP-1, and p62 KD MDA-MB-231 cells, exhibit enhanced autophagy activation in response to TLR4 stimulation. ( A ) p62 KD THP-1 cells were generated, and the knockdown efficacy of p62 was confirmed with anti-p62 antibody. ( B , C ) Ctrl and p62 KD THP-1 cells were treated with or without vehicle or CQ (10 μM), in the presence or absence of LPS (10 μg/mL), for 6 h. Whole cell lysates were immunoblotted with anti-LC3A/B antibody and anti-GAPDH antibody as a loading control ( B ). The LC3II levels were analyzed with Image J program ( C ). Data shown are averages from a minimum of 3 independent experiments (± SEM). *, p

    Article Snippet: Human hepatic adenocarcinoma cell line SK-HEP-1 (ATCC, HTB-52), human breast adenocarcinoma cell line MDA-MB-231 (ATCC, HTB-26), and human lung cancer cell line A549 (ATCC, CCL-185) were purchased from ATCC, and cultured in DMEM or RPMI contained with 10% FBS.

    Techniques: Multiple Displacement Amplification, Activation Assay, Generated

    c-MYC associated with PCAF and CtBPs to assemble a complex. (A) The Flag-c-MYC-associated complex. The pCDNA3-2×Flag (empty vector, EV) and pCDNA3-2×Flag-c-MYC plasmids were transfected into MIA PaCa-2 cells, respectively. The resulting cells were subjected to immunoprecipitation with the anti-Flag resin. The purified complexes were separated in an SDS-PAGE gel and incubated with a silver staining kit. The IgG and Flag-c-MYC were indicated by arrows. (B) c-MYC could pull down PCAF and CtBPs in vivo . Equal weight of pancreatic tissues from three AP patients was mixed and lysed, and 1/11 cell extracts were used as an input, and the other 10/11 cell extracts were equally divided into two parts, followed by immunoprecipitation with an IgG and anti-c-MYC antibody-associated protein A beads, respectively. The input and output proteins were used to determine protein levels of c-MYC, PCAF, CtBP1 and CtBP2, respectively. (C) c-MYC directly interacted with PCAF but not CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-PCAF, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-PCAF. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies. (D) PCAF directly interacted with both c-MYC and CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output protein levels using anti-Flag and anti-Myc antibodies. (E) CtBPs assembled a heterotetramer in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies.

    Journal: International Journal of Biological Sciences

    Article Title: Inflammation and DNA methylation coregulate the CtBP-PCAF-c-MYC transcriptional complex to activate the expression of a long non-coding RNA CASC2 in acute pancreatitis

    doi: 10.7150/ijbs.43557

    Figure Lengend Snippet: c-MYC associated with PCAF and CtBPs to assemble a complex. (A) The Flag-c-MYC-associated complex. The pCDNA3-2×Flag (empty vector, EV) and pCDNA3-2×Flag-c-MYC plasmids were transfected into MIA PaCa-2 cells, respectively. The resulting cells were subjected to immunoprecipitation with the anti-Flag resin. The purified complexes were separated in an SDS-PAGE gel and incubated with a silver staining kit. The IgG and Flag-c-MYC were indicated by arrows. (B) c-MYC could pull down PCAF and CtBPs in vivo . Equal weight of pancreatic tissues from three AP patients was mixed and lysed, and 1/11 cell extracts were used as an input, and the other 10/11 cell extracts were equally divided into two parts, followed by immunoprecipitation with an IgG and anti-c-MYC antibody-associated protein A beads, respectively. The input and output proteins were used to determine protein levels of c-MYC, PCAF, CtBP1 and CtBP2, respectively. (C) c-MYC directly interacted with PCAF but not CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-PCAF, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-c-MYC + pCDNA3-6×Myc-PCAF. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies. (D) PCAF directly interacted with both c-MYC and CtBPs in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-PCAF + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output protein levels using anti-Flag and anti-Myc antibodies. (E) CtBPs assembled a heterotetramer in vitro . The MIA PaCa-2 cells were transfected with different plasmids including pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag + pCDNA3-6×Myc-CtBP2, pCDNA3-2×Flag + pCDNA3-6×Myc-c-MYC, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP1, pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-CtBP2, and pCDNA3-2×Flag-CtBP1 + pCDNA3-6×Myc-c-MYC. The resulting cells were lysed and immunoprecipitated with an anti-Flag and anti-Myc resins, respectively, followed by immunoblots to examine the input and output proteins levels using anti-Flag and anti-Myc antibodies.

    Article Snippet: Cell culture and transfectionThe human pancreatic epithelial cell line MIA PaCa-2 was obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA, #CRL-1420) and grown in DMEM (Dulbecco's modified Eagle's medium) (Sigma-Aldrich, #D6046) supplemented with 10% FBS (Fetal Bovine Serum) (Thermo Fisher Scientific, #16000044) and 1% PS (Penicillin-Streptomycin) (Thermo Fisher Scientific, #15140163).

    Techniques: Plasmid Preparation, Transfection, Immunoprecipitation, Purification, SDS Page, Incubation, Silver Staining, In Vivo, In Vitro, Western Blot

    Recombinant IL6 and TNF-α induced the expression of CtBPs at both transcriptional and protein levels. (A) IL6 and TNF-α induced the mRNA levels of CtBPs . The MIA PaCa-2 cells were treated with IL6 and TNF-α at the concentrations of 0, 10, 25 and 50 ng/mL for 6 h. The resulting cells were used for RNA isolation, followed by qRT-PCR analyses to determine the mRNA levels of CtBP1 and CtBP2 . ** P

    Journal: International Journal of Biological Sciences

    Article Title: Inflammation and DNA methylation coregulate the CtBP-PCAF-c-MYC transcriptional complex to activate the expression of a long non-coding RNA CASC2 in acute pancreatitis

    doi: 10.7150/ijbs.43557

    Figure Lengend Snippet: Recombinant IL6 and TNF-α induced the expression of CtBPs at both transcriptional and protein levels. (A) IL6 and TNF-α induced the mRNA levels of CtBPs . The MIA PaCa-2 cells were treated with IL6 and TNF-α at the concentrations of 0, 10, 25 and 50 ng/mL for 6 h. The resulting cells were used for RNA isolation, followed by qRT-PCR analyses to determine the mRNA levels of CtBP1 and CtBP2 . ** P

    Article Snippet: Cell culture and transfectionThe human pancreatic epithelial cell line MIA PaCa-2 was obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA, #CRL-1420) and grown in DMEM (Dulbecco's modified Eagle's medium) (Sigma-Aldrich, #D6046) supplemented with 10% FBS (Fetal Bovine Serum) (Thermo Fisher Scientific, #16000044) and 1% PS (Penicillin-Streptomycin) (Thermo Fisher Scientific, #15140163).

    Techniques: Recombinant, Expressing, Isolation, Quantitative RT-PCR

    Continuous C. pneumoniae infection in HEp-2 cells inoculated with isolate CM-1 (A to D) or isolate TW-183 (E and F). (A and B) Typical inclusions; (C to F) altered inclusions; (D) aberrant inclusion. C, cytoplasm; N, nucleus; M, mitochondria; im, inclusion membrane; e, intrainclusional membranous material.

    Journal: Journal of Clinical Microbiology

    Article Title: Ultrastructural Study of Chlamydia pneumoniae In a Continuous-Infection Model

    doi: 10.1128/JCM.39.10.3721-3723.2001

    Figure Lengend Snippet: Continuous C. pneumoniae infection in HEp-2 cells inoculated with isolate CM-1 (A to D) or isolate TW-183 (E and F). (A and B) Typical inclusions; (C to F) altered inclusions; (D) aberrant inclusion. C, cytoplasm; N, nucleus; M, mitochondria; im, inclusion membrane; e, intrainclusional membranous material.

    Article Snippet: Briefly, confluent HEp-2 cells were inoculated once with C. pneumoniae isolate TW-183 (ATCC VR2282) or CM-1 (ATCC VR1360) to achieve 100% infection.

    Techniques: Infection