vk2 e6e7 cells  (Millipore)


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    Structured Review

    Millipore vk2 e6e7 cells
    Immunofluroscence of phosphorylated p65-NF-κB expression in hPVECs and <t>VK2/E6E7</t> cells. hPVECs (A) and VK2/E6E7 (B) cells were treated with LPS (10 μg/ml 6 hrs) or Bay 11–7082 (5 μM for 24 hrs). Stimulation with LPS activates p65-NF-κB expression in hPVECs (b,e) and VK2/E6E7 cells (h,k) as compared to unstimulated hPVECs (a,d) and VK2/E6E7 cells (g,j). Treatment with Bay 11–7082 attenuated NF-κB expression in hPVECs (c,f) and VK2/E6E7 cells (i,l). Nucleus was stained with DAPI (blue), p65-NF-κB was stained with FITC (green), FITC and DAPI merged (d,e,f,j,k,l). The images shown are the representative pictures of one of three identical experiments performed on three different days (Mag. X 63).
    Vk2 E6e7 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vk2 e6e7 cells - by Bioz Stars, 2020-08
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    Images

    1) Product Images from "Expression of hemoglobin-α and β subunits in human vaginal epithelial cells and their functional significance"

    Article Title: Expression of hemoglobin-α and β subunits in human vaginal epithelial cells and their functional significance

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0171084

    Immunofluroscence of phosphorylated p65-NF-κB expression in hPVECs and VK2/E6E7 cells. hPVECs (A) and VK2/E6E7 (B) cells were treated with LPS (10 μg/ml 6 hrs) or Bay 11–7082 (5 μM for 24 hrs). Stimulation with LPS activates p65-NF-κB expression in hPVECs (b,e) and VK2/E6E7 cells (h,k) as compared to unstimulated hPVECs (a,d) and VK2/E6E7 cells (g,j). Treatment with Bay 11–7082 attenuated NF-κB expression in hPVECs (c,f) and VK2/E6E7 cells (i,l). Nucleus was stained with DAPI (blue), p65-NF-κB was stained with FITC (green), FITC and DAPI merged (d,e,f,j,k,l). The images shown are the representative pictures of one of three identical experiments performed on three different days (Mag. X 63).
    Figure Legend Snippet: Immunofluroscence of phosphorylated p65-NF-κB expression in hPVECs and VK2/E6E7 cells. hPVECs (A) and VK2/E6E7 (B) cells were treated with LPS (10 μg/ml 6 hrs) or Bay 11–7082 (5 μM for 24 hrs). Stimulation with LPS activates p65-NF-κB expression in hPVECs (b,e) and VK2/E6E7 cells (h,k) as compared to unstimulated hPVECs (a,d) and VK2/E6E7 cells (g,j). Treatment with Bay 11–7082 attenuated NF-κB expression in hPVECs (c,f) and VK2/E6E7 cells (i,l). Nucleus was stained with DAPI (blue), p65-NF-κB was stained with FITC (green), FITC and DAPI merged (d,e,f,j,k,l). The images shown are the representative pictures of one of three identical experiments performed on three different days (Mag. X 63).

    Techniques Used: Expressing, Staining

    Cultures of human primary vaginal epithelial cells (hPVECS) and VK2/E6E7 cells. Different phases of growing cells of hPVECs are shown (a-d). a: Cells attached to the surface of the culture flask on day 10 (10x), b: on day 15 (40x), c: confluent cells on day 25 (10x) and d: confluent cells on day 25 (40x); e, f, g: confluent hPVECs from three different patient samples; h: confluent cultures of VK2/E6E7 cell line on day 10 (40x).
    Figure Legend Snippet: Cultures of human primary vaginal epithelial cells (hPVECS) and VK2/E6E7 cells. Different phases of growing cells of hPVECs are shown (a-d). a: Cells attached to the surface of the culture flask on day 10 (10x), b: on day 15 (40x), c: confluent cells on day 25 (10x) and d: confluent cells on day 25 (40x); e, f, g: confluent hPVECs from three different patient samples; h: confluent cultures of VK2/E6E7 cell line on day 10 (40x).

    Techniques Used:

    Immunofluorescence localization of cytokeratin-13. Confocal images showing cytoplasmic localization of cytokeratin-13(green) in hPVECs (a-c) and VK2/E6E7 cells (d-f). Nucleus was stained with DAPI (blue). FITC (a, d), FITC and DAPI merge (b,e) and no primary antibody controls (c,f) are shown. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).
    Figure Legend Snippet: Immunofluorescence localization of cytokeratin-13. Confocal images showing cytoplasmic localization of cytokeratin-13(green) in hPVECs (a-c) and VK2/E6E7 cells (d-f). Nucleus was stained with DAPI (blue). FITC (a, d), FITC and DAPI merge (b,e) and no primary antibody controls (c,f) are shown. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).

    Techniques Used: Immunofluorescence, Staining

    TLR4 expression in hPVECs and VK2/E6E7 cells. (A). RT-PCR analysis of TLR4 expression in VK2/E6E7 cells and hPVECs. expression of TLR4 (182 bp) in VK2/E6E7 cells (a), (c), and hPVECs (b),(d) was observed. 1: Untreated cells (control); 2: DMSO treated cells (control); 3: TLR4 antibody treated cells; 4: LPS treated cells (10 μg/ml for 6 hrs) and 5: Bay 11–7082 treated cells (5μM for 24 hrs) stimulated with LPS (10μg/ml for 6 hrs) and (c), (d) Housekeeping gene Gapdh (238 bp) considered as an internal standard. The gel picture shown is one of the representative pictures from three independent experiments. (B). Densitometric analysis of bands from RT–PCR amplification products of TLR4 mRNA reported in Fig 10A. The expression of TLR4 mRNA was up-regulated in LPS-induced cells. Bay 11–7082 treatment has no effect on the expression of TLR4 mRNA in cells induced with LPS.
    Figure Legend Snippet: TLR4 expression in hPVECs and VK2/E6E7 cells. (A). RT-PCR analysis of TLR4 expression in VK2/E6E7 cells and hPVECs. expression of TLR4 (182 bp) in VK2/E6E7 cells (a), (c), and hPVECs (b),(d) was observed. 1: Untreated cells (control); 2: DMSO treated cells (control); 3: TLR4 antibody treated cells; 4: LPS treated cells (10 μg/ml for 6 hrs) and 5: Bay 11–7082 treated cells (5μM for 24 hrs) stimulated with LPS (10μg/ml for 6 hrs) and (c), (d) Housekeeping gene Gapdh (238 bp) considered as an internal standard. The gel picture shown is one of the representative pictures from three independent experiments. (B). Densitometric analysis of bands from RT–PCR amplification products of TLR4 mRNA reported in Fig 10A. The expression of TLR4 mRNA was up-regulated in LPS-induced cells. Bay 11–7082 treatment has no effect on the expression of TLR4 mRNA in cells induced with LPS.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. (A) RT-PCR analysis; 1: Untreated hPVECs, 2: hPVECs treated for 6 hrs with LPS (10 μg/ml), 3: Untreated VK2/E6E7 cells and 4: VK2 cells treated with LPS (10 μg/ml) for 6 hrs. Loading control, Gapdh (238 bp) expression in hPVECs. The gels shown are one of the representative pictures from three independent experiments performed on three different days. (B) Densitometric analysis of bands from RT-PCR amplification products of Hb-α and Hb-β mRNAs shown in figure-5A.
    Figure Legend Snippet: Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. (A) RT-PCR analysis; 1: Untreated hPVECs, 2: hPVECs treated for 6 hrs with LPS (10 μg/ml), 3: Untreated VK2/E6E7 cells and 4: VK2 cells treated with LPS (10 μg/ml) for 6 hrs. Loading control, Gapdh (238 bp) expression in hPVECs. The gels shown are one of the representative pictures from three independent experiments performed on three different days. (B) Densitometric analysis of bands from RT-PCR amplification products of Hb-α and Hb-β mRNAs shown in figure-5A.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    Immunofluorescence localization of vimentin. hPVECs (a-c) and VK2/E6E7 cells (d-f) did not express vimentin. Cytoplasmic localization of vimentin (green) is seen in HeLa cells (positive control) (g-i). Nucleus was stained with DAPI (blue), FITC (a, d, g), FITC and DAPI merge (b,e,h) and no primary antibody controls (c,f,i) are indicated. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).
    Figure Legend Snippet: Immunofluorescence localization of vimentin. hPVECs (a-c) and VK2/E6E7 cells (d-f) did not express vimentin. Cytoplasmic localization of vimentin (green) is seen in HeLa cells (positive control) (g-i). Nucleus was stained with DAPI (blue), FITC (a, d, g), FITC and DAPI merge (b,e,h) and no primary antibody controls (c,f,i) are indicated. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).

    Techniques Used: Immunofluorescence, Positive Control, Staining

    Expression of NF- κ B in hPVECs and VK2/E6E7 cells. (A). NF-κB levels in un-stimulated, LPS stimulated and Bay 11–7082 treated hPVECs and VK2/E6E7 cells analyzed by ELISA. Cells were seeded at a density of 10 6 /well in 24-well plates and induced with LPS (10 μg/ml for 6 hrs) or Bay-11-0782 (5 μM for 24 hrs). Levels of p65-NF-κB were up-regulated in LPS-induced cells, where as Bay-11-7082 reversed this effect (a). Values represent mean ± SD of three experiments performed on different days. Values are statistically significant (***p
    Figure Legend Snippet: Expression of NF- κ B in hPVECs and VK2/E6E7 cells. (A). NF-κB levels in un-stimulated, LPS stimulated and Bay 11–7082 treated hPVECs and VK2/E6E7 cells analyzed by ELISA. Cells were seeded at a density of 10 6 /well in 24-well plates and induced with LPS (10 μg/ml for 6 hrs) or Bay-11-0782 (5 μM for 24 hrs). Levels of p65-NF-κB were up-regulated in LPS-induced cells, where as Bay-11-7082 reversed this effect (a). Values represent mean ± SD of three experiments performed on different days. Values are statistically significant (***p

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay

    RT-PCR analysis of Hb-α and Hb-β in presence of LPS and Bay 11–7082. (A) Hb-α (429 bp) (a) and Hb-β (444 bp) (b) expression was observed in hPVECs (1,2) and VK2/E6E7 cells (3,4) before and after the treatment with LPS (10 μg/ml for 6 hrs) and Bay 11–7082 (5μM for 24 hrs). LPS-induced (a1, a3, b1, b3), Bay 11–7082 treated (a2, a4, b2, b4) are shown. Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells, whereas Bay 11–7082 attenuated this expression (c) Gapdh (238 bp) was used as loading control. The gels shown are the representative pictures from three independent experiments. (B). Densitometric analysis of RT-PCR bands shown in Fig 12A. Expression of both Hb-α and Hb-β was elevated in LPS-stimulated cells. In contrast, Bay 11–7082 attenuated the expression of Hb-α and Hb-β .
    Figure Legend Snippet: RT-PCR analysis of Hb-α and Hb-β in presence of LPS and Bay 11–7082. (A) Hb-α (429 bp) (a) and Hb-β (444 bp) (b) expression was observed in hPVECs (1,2) and VK2/E6E7 cells (3,4) before and after the treatment with LPS (10 μg/ml for 6 hrs) and Bay 11–7082 (5μM for 24 hrs). LPS-induced (a1, a3, b1, b3), Bay 11–7082 treated (a2, a4, b2, b4) are shown. Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells, whereas Bay 11–7082 attenuated this expression (c) Gapdh (238 bp) was used as loading control. The gels shown are the representative pictures from three independent experiments. (B). Densitometric analysis of RT-PCR bands shown in Fig 12A. Expression of both Hb-α and Hb-β was elevated in LPS-stimulated cells. In contrast, Bay 11–7082 attenuated the expression of Hb-α and Hb-β .

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    Western blot analysis of biomarker expression in hPVECs, VK2/E6E7 cells and RBCs. (a) Cytokeratin-13 (47kDa) expression: Lane-1: RBCs, lane-2: hPVECs and lane-3: VK2/E6E7 cells. (b) Solute carrier family 4 member-1 (SLC4A1) (100 kDa) expression: Lane-1: RBCs (positive control), lane-2: hPVECs, lane-3: VK2/E6E7 cells. (c)Vimentin (54 kDa) expression: Lane-1: vimentin expression in HeLa cells (positive control), Lane-2: hPVECs, Lane-3: VK2/E6E7 cells. (d) β-actin (42 kDa) expression in hPVECs (loading control). The blots shown are one of the representative pictures from three independent experiments performed on three different days.
    Figure Legend Snippet: Western blot analysis of biomarker expression in hPVECs, VK2/E6E7 cells and RBCs. (a) Cytokeratin-13 (47kDa) expression: Lane-1: RBCs, lane-2: hPVECs and lane-3: VK2/E6E7 cells. (b) Solute carrier family 4 member-1 (SLC4A1) (100 kDa) expression: Lane-1: RBCs (positive control), lane-2: hPVECs, lane-3: VK2/E6E7 cells. (c)Vimentin (54 kDa) expression: Lane-1: vimentin expression in HeLa cells (positive control), Lane-2: hPVECs, Lane-3: VK2/E6E7 cells. (d) β-actin (42 kDa) expression in hPVECs (loading control). The blots shown are one of the representative pictures from three independent experiments performed on three different days.

    Techniques Used: Western Blot, Biomarker Assay, Expressing, Positive Control

    RT-PCR analysis of human-β defensin-1 ( hBD-1 ). (A) mRNA expression of hBD1 (139bp) was observed in un-stimulated (a1, a3), LPS stimulated (a2, a4) in hPVECs (a1, a2) and VK2/E6E7 cells (a3, a4). Cells were seeded at a density of 10 6 /well in a 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Expression of hBD1 mRNA was up-regulated in LPS-induced cells. Representative image of RT-PCR analysis of hBD-1 mRNAs expression is shown. Gapdh confirmed roughly equivalent loading of RNA samples. 1: hPVECs (Unstimulated); 2:hPVECs induced with LPS; 3:VK2/E6E7 cells (Unstimulated); 4:VK2/E6E7 cells stimulated with LPS (10μg/ml). (B). Densitometric analysis of bands from RT-PCR amplification products of hBD1 mRNA reported in Figure 13A. Expression of hBD-1 was increased in LPS-stimulated hPVECS and VK2/E6E7 cells.
    Figure Legend Snippet: RT-PCR analysis of human-β defensin-1 ( hBD-1 ). (A) mRNA expression of hBD1 (139bp) was observed in un-stimulated (a1, a3), LPS stimulated (a2, a4) in hPVECs (a1, a2) and VK2/E6E7 cells (a3, a4). Cells were seeded at a density of 10 6 /well in a 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Expression of hBD1 mRNA was up-regulated in LPS-induced cells. Representative image of RT-PCR analysis of hBD-1 mRNAs expression is shown. Gapdh confirmed roughly equivalent loading of RNA samples. 1: hPVECs (Unstimulated); 2:hPVECs induced with LPS; 3:VK2/E6E7 cells (Unstimulated); 4:VK2/E6E7 cells stimulated with LPS (10μg/ml). (B). Densitometric analysis of bands from RT-PCR amplification products of hBD1 mRNA reported in Figure 13A. Expression of hBD-1 was increased in LPS-stimulated hPVECS and VK2/E6E7 cells.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Amplification

    Cytokoine ELISA of hPVECs and VK2/E6E7 cells. IL-6 (A) and IL-10 (B) in the culture supernatants of un-stimulated and LPS stimulated hPVECs and VK2/E6E7 cells at indicated time points (6, 12, 24 hrs). Cells were seeded at a density of 10 6 /well in 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Levels of IL-6 were up-regulated in LPS-induced cells. Values represent the mean ± SD of three independent experiments performed on different days. Level of significance (*p
    Figure Legend Snippet: Cytokoine ELISA of hPVECs and VK2/E6E7 cells. IL-6 (A) and IL-10 (B) in the culture supernatants of un-stimulated and LPS stimulated hPVECs and VK2/E6E7 cells at indicated time points (6, 12, 24 hrs). Cells were seeded at a density of 10 6 /well in 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Levels of IL-6 were up-regulated in LPS-induced cells. Values represent the mean ± SD of three independent experiments performed on different days. Level of significance (*p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells at protein level. (A, B). Immunofluorescence localization of Hb-α and Hb-β. Cytoplasmic localization (green) of Hb-α (A) and Hb-β (B) was observed in hPVECs (a,b,c) and VK2/E6E7 cells (d,e,f). DAPI stained nucleus (blue). a, d: Untreated cells; b,e: LPS treated cells; c,f: Primary antibody controls did not show Hb-α and Hb-β localization. Expression was significantly up-regulated when hPVECs and VK2/E6E7 cells stimulated with LPS (10 μg/ml for 6 hrs) (b, e). The figure shown is one of the representative pictures from three independent experiments performed on three different days (Mag. 63X). (C). Western blot analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. β-actin (42 kDa) (loading control) expression in hPVECs (1: Untreated hPVECs, 2: hPVECs induced with LPS, 3: Untreated VK2/E6E7 cells and 4: VK2/E6E7 cells induced with LPS). (D). Densitometric analysis of bands from western blot of Hb-α and Hb-β shown in Fig 7C.
    Figure Legend Snippet: Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells at protein level. (A, B). Immunofluorescence localization of Hb-α and Hb-β. Cytoplasmic localization (green) of Hb-α (A) and Hb-β (B) was observed in hPVECs (a,b,c) and VK2/E6E7 cells (d,e,f). DAPI stained nucleus (blue). a, d: Untreated cells; b,e: LPS treated cells; c,f: Primary antibody controls did not show Hb-α and Hb-β localization. Expression was significantly up-regulated when hPVECs and VK2/E6E7 cells stimulated with LPS (10 μg/ml for 6 hrs) (b, e). The figure shown is one of the representative pictures from three independent experiments performed on three different days (Mag. 63X). (C). Western blot analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. β-actin (42 kDa) (loading control) expression in hPVECs (1: Untreated hPVECs, 2: hPVECs induced with LPS, 3: Untreated VK2/E6E7 cells and 4: VK2/E6E7 cells induced with LPS). (D). Densitometric analysis of bands from western blot of Hb-α and Hb-β shown in Fig 7C.

    Techniques Used: Expressing, Immunofluorescence, Staining, Western Blot

    qPCR analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells before and after induction with LPS. Cells seeded at a density of 10 6 /well in 24-well plates were treated with LPS (10 μg/ml for 6 hrs). Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells. Each bar represents the mean ± SD of three independent replicates (***p
    Figure Legend Snippet: qPCR analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells before and after induction with LPS. Cells seeded at a density of 10 6 /well in 24-well plates were treated with LPS (10 μg/ml for 6 hrs). Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells. Each bar represents the mean ± SD of three independent replicates (***p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    2) Product Images from "Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection"

    Article Title: Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0114208

    Subcellular location of intracellular bacteria in mitosis. A. VK2/E6E7 cells were infected with DyLight 594 NHS Ester-stained N. gonorrhoeae . Cells were maintained in a live-cell incubator connected to an inverted fluorescence microscope. Cellular DNA was stained with Hoechst 33342 and is shown in blue. Images of live mitotic cells were taken through a 100x oil objective in 40–60 z-stacks. Images were further deconvolved and processed by Inside 4D software. Bacteria (red) present during prophase (upper), prometaphase (mid) and anaphase (lower) are shown. Bacteria that are in close proximity to the chromatin are marked with arrows. The scale bar represents 10 µm. B. Viable bacteria associate with chromatin in ELISA. Chromatin, extracted from 1.25×10 5 cells was coated in each well. Viable (5×10 6 CFU/well or 1×10 6 CFU/well) or PFA-fixed (5×10 6 CFU/well) MS11P+ was allowed to interact with the chromatin for 2 h at 37°C and 5% CO 2 . Rabbit anti- N. gonorrhoeae antibodies followed by goat anti rabbit IgG-HRP antibodies were used to detect the bacteria bound to the chromatin. Absorbance was read at 450 nm. Background was measured in wells containing all agents except chromatin. Shown is a graph of one assay in technical triplicate and standard deviations.
    Figure Legend Snippet: Subcellular location of intracellular bacteria in mitosis. A. VK2/E6E7 cells were infected with DyLight 594 NHS Ester-stained N. gonorrhoeae . Cells were maintained in a live-cell incubator connected to an inverted fluorescence microscope. Cellular DNA was stained with Hoechst 33342 and is shown in blue. Images of live mitotic cells were taken through a 100x oil objective in 40–60 z-stacks. Images were further deconvolved and processed by Inside 4D software. Bacteria (red) present during prophase (upper), prometaphase (mid) and anaphase (lower) are shown. Bacteria that are in close proximity to the chromatin are marked with arrows. The scale bar represents 10 µm. B. Viable bacteria associate with chromatin in ELISA. Chromatin, extracted from 1.25×10 5 cells was coated in each well. Viable (5×10 6 CFU/well or 1×10 6 CFU/well) or PFA-fixed (5×10 6 CFU/well) MS11P+ was allowed to interact with the chromatin for 2 h at 37°C and 5% CO 2 . Rabbit anti- N. gonorrhoeae antibodies followed by goat anti rabbit IgG-HRP antibodies were used to detect the bacteria bound to the chromatin. Absorbance was read at 450 nm. Background was measured in wells containing all agents except chromatin. Shown is a graph of one assay in technical triplicate and standard deviations.

    Techniques Used: Infection, Staining, Fluorescence, Microscopy, Software, Enzyme-linked Immunosorbent Assay

    Gonococcal infection causes durations of mitosis, nuclear swelling, and targets regulatory mitotic genes and proteins. A. VK2/E6E7 cells were synchronized and bacteria were allowed to adhere and invade cells for 24 h. Cells were then fixed and stained with DAPI. The nuclear area of 300 infected and 300 uninfected cells were measured. Data were analyzed using a paired 2-tailed Student's t -test. The average nuclear areas observed in 3 independent experiments are shown (* p
    Figure Legend Snippet: Gonococcal infection causes durations of mitosis, nuclear swelling, and targets regulatory mitotic genes and proteins. A. VK2/E6E7 cells were synchronized and bacteria were allowed to adhere and invade cells for 24 h. Cells were then fixed and stained with DAPI. The nuclear area of 300 infected and 300 uninfected cells were measured. Data were analyzed using a paired 2-tailed Student's t -test. The average nuclear areas observed in 3 independent experiments are shown (* p

    Techniques Used: Infection, Staining

    Bacterial infection induces micronuclei formation in VK2/E6E7 cells. VK2/E6E7 cells were infected with N. gonorrhoeae for 24 h. Cytokinesis was blocked with cytochalasin B for 36 h and BNC from infected and control cells were analyzed for micronuclei formation. A. Average numbers of observed micronuclei/1000 BNC ± standard deviation from 3 independent experiments are shown (* p
    Figure Legend Snippet: Bacterial infection induces micronuclei formation in VK2/E6E7 cells. VK2/E6E7 cells were infected with N. gonorrhoeae for 24 h. Cytokinesis was blocked with cytochalasin B for 36 h and BNC from infected and control cells were analyzed for micronuclei formation. A. Average numbers of observed micronuclei/1000 BNC ± standard deviation from 3 independent experiments are shown (* p

    Techniques Used: Infection, Standard Deviation

    Microinjection of bacterial lysates in the cytoplasm of VK2/E6E7 cells causes DSBs. A. Interphase VK2/E6E7 cells were subjected to cytoplasmic microinjection of bacterial MS11 P+ lysate, MS11 P+ HI lysate, or PBS. Cells were incubated for 20–24 h and then stained for DSBs with 53BP1 antibodies. The graph shows the average number of 53BP1 positive cells counted in two independent experiments under each condition. Control cells are non-injected cells. B. Images showing DIC and fluorescent images of representative cells microinjected with MS11 P+ lysate (left) or PBS (right). FITC-dextran (green) was co-injected into the cytoplasm to identify microinjected cells. Scale bar represents 10 µm.
    Figure Legend Snippet: Microinjection of bacterial lysates in the cytoplasm of VK2/E6E7 cells causes DSBs. A. Interphase VK2/E6E7 cells were subjected to cytoplasmic microinjection of bacterial MS11 P+ lysate, MS11 P+ HI lysate, or PBS. Cells were incubated for 20–24 h and then stained for DSBs with 53BP1 antibodies. The graph shows the average number of 53BP1 positive cells counted in two independent experiments under each condition. Control cells are non-injected cells. B. Images showing DIC and fluorescent images of representative cells microinjected with MS11 P+ lysate (left) or PBS (right). FITC-dextran (green) was co-injected into the cytoplasm to identify microinjected cells. Scale bar represents 10 µm.

    Techniques Used: Incubation, Staining, Injection

    Lysates of N. gonorrhoeae fragments pECFP-N1 and damage DNA from VK2/E6E7 cells. A. DNA agarose gel showing the digestion of pECFP-N1 plasmid by HindIII (positive control, lane 2), MS11 P+ lysate (lane 3), and MS11 P+ HI lysate (lane 5). Lane 5 shows bacterial MS11 P+ lysate without pECFP-N1 and lane 1 shows uncut circular pECFP-N1. B. PFGE analysis of purified VK2/E6E7 genomic DNA treated for 24 h with: lane 1: PBS (negative control), lane 2: MS11 P+ lysate, lane 3: MS11 P+ HI lysate. Lane 4 shows bacterial MS11 P+ lysate without VK2/E6E7 genomic DNA. C. Graph showing quantification of DNA smears (measured directly underneath and below the band). Shown are smear pixel intensities of cellular DNA alone and cellular DNA exposed to bacterial lysates and HI bacterial lysates. D. PFGE showing genomic DNA subjected to commercial restriction enzymes for 24 h. Lane 1: DNA incubated with CutSmart reaction buffer (negative control). Lane 2: DNA incubated with NgoMIV. Lane 3: DNA incubated with MfeI, Lane 4: DNA incubated with NgoMIV and MfeI Lane 5: DNA incubated with NgoMIV and BamHI/KpnI/MfeI (BKM).
    Figure Legend Snippet: Lysates of N. gonorrhoeae fragments pECFP-N1 and damage DNA from VK2/E6E7 cells. A. DNA agarose gel showing the digestion of pECFP-N1 plasmid by HindIII (positive control, lane 2), MS11 P+ lysate (lane 3), and MS11 P+ HI lysate (lane 5). Lane 5 shows bacterial MS11 P+ lysate without pECFP-N1 and lane 1 shows uncut circular pECFP-N1. B. PFGE analysis of purified VK2/E6E7 genomic DNA treated for 24 h with: lane 1: PBS (negative control), lane 2: MS11 P+ lysate, lane 3: MS11 P+ HI lysate. Lane 4 shows bacterial MS11 P+ lysate without VK2/E6E7 genomic DNA. C. Graph showing quantification of DNA smears (measured directly underneath and below the band). Shown are smear pixel intensities of cellular DNA alone and cellular DNA exposed to bacterial lysates and HI bacterial lysates. D. PFGE showing genomic DNA subjected to commercial restriction enzymes for 24 h. Lane 1: DNA incubated with CutSmart reaction buffer (negative control). Lane 2: DNA incubated with NgoMIV. Lane 3: DNA incubated with MfeI, Lane 4: DNA incubated with NgoMIV and MfeI Lane 5: DNA incubated with NgoMIV and BamHI/KpnI/MfeI (BKM).

    Techniques Used: Agarose Gel Electrophoresis, Plasmid Preparation, Positive Control, Purification, Negative Control, Incubation

    3) Product Images from "Expression of hemoglobin-α and β subunits in human vaginal epithelial cells and their functional significance"

    Article Title: Expression of hemoglobin-α and β subunits in human vaginal epithelial cells and their functional significance

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0171084

    Immunofluroscence of phosphorylated p65-NF-κB expression in hPVECs and VK2/E6E7 cells. hPVECs (A) and VK2/E6E7 (B) cells were treated with LPS (10 μg/ml 6 hrs) or Bay 11–7082 (5 μM for 24 hrs). Stimulation with LPS activates p65-NF-κB expression in hPVECs (b,e) and VK2/E6E7 cells (h,k) as compared to unstimulated hPVECs (a,d) and VK2/E6E7 cells (g,j). Treatment with Bay 11–7082 attenuated NF-κB expression in hPVECs (c,f) and VK2/E6E7 cells (i,l). Nucleus was stained with DAPI (blue), p65-NF-κB was stained with FITC (green), FITC and DAPI merged (d,e,f,j,k,l). The images shown are the representative pictures of one of three identical experiments performed on three different days (Mag. X 63).
    Figure Legend Snippet: Immunofluroscence of phosphorylated p65-NF-κB expression in hPVECs and VK2/E6E7 cells. hPVECs (A) and VK2/E6E7 (B) cells were treated with LPS (10 μg/ml 6 hrs) or Bay 11–7082 (5 μM for 24 hrs). Stimulation with LPS activates p65-NF-κB expression in hPVECs (b,e) and VK2/E6E7 cells (h,k) as compared to unstimulated hPVECs (a,d) and VK2/E6E7 cells (g,j). Treatment with Bay 11–7082 attenuated NF-κB expression in hPVECs (c,f) and VK2/E6E7 cells (i,l). Nucleus was stained with DAPI (blue), p65-NF-κB was stained with FITC (green), FITC and DAPI merged (d,e,f,j,k,l). The images shown are the representative pictures of one of three identical experiments performed on three different days (Mag. X 63).

    Techniques Used: Expressing, Staining

    Cultures of human primary vaginal epithelial cells (hPVECS) and VK2/E6E7 cells. Different phases of growing cells of hPVECs are shown (a-d). a: Cells attached to the surface of the culture flask on day 10 (10x), b: on day 15 (40x), c: confluent cells on day 25 (10x) and d: confluent cells on day 25 (40x); e, f, g: confluent hPVECs from three different patient samples; h: confluent cultures of VK2/E6E7 cell line on day 10 (40x).
    Figure Legend Snippet: Cultures of human primary vaginal epithelial cells (hPVECS) and VK2/E6E7 cells. Different phases of growing cells of hPVECs are shown (a-d). a: Cells attached to the surface of the culture flask on day 10 (10x), b: on day 15 (40x), c: confluent cells on day 25 (10x) and d: confluent cells on day 25 (40x); e, f, g: confluent hPVECs from three different patient samples; h: confluent cultures of VK2/E6E7 cell line on day 10 (40x).

    Techniques Used:

    Immunofluorescence localization of cytokeratin-13. Confocal images showing cytoplasmic localization of cytokeratin-13(green) in hPVECs (a-c) and VK2/E6E7 cells (d-f). Nucleus was stained with DAPI (blue). FITC (a, d), FITC and DAPI merge (b,e) and no primary antibody controls (c,f) are shown. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).
    Figure Legend Snippet: Immunofluorescence localization of cytokeratin-13. Confocal images showing cytoplasmic localization of cytokeratin-13(green) in hPVECs (a-c) and VK2/E6E7 cells (d-f). Nucleus was stained with DAPI (blue). FITC (a, d), FITC and DAPI merge (b,e) and no primary antibody controls (c,f) are shown. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).

    Techniques Used: Immunofluorescence, Staining

    TLR4 expression in hPVECs and VK2/E6E7 cells. (A). RT-PCR analysis of TLR4 expression in VK2/E6E7 cells and hPVECs. expression of TLR4 (182 bp) in VK2/E6E7 cells (a), (c), and hPVECs (b),(d) was observed. 1: Untreated cells (control); 2: DMSO treated cells (control); 3: TLR4 antibody treated cells; 4: LPS treated cells (10 μg/ml for 6 hrs) and 5: Bay 11–7082 treated cells (5μM for 24 hrs) stimulated with LPS (10μg/ml for 6 hrs) and (c), (d) Housekeeping gene Gapdh (238 bp) considered as an internal standard. The gel picture shown is one of the representative pictures from three independent experiments. (B). Densitometric analysis of bands from RT–PCR amplification products of TLR4 mRNA reported in Fig 10A. The expression of TLR4 mRNA was up-regulated in LPS-induced cells. Bay 11–7082 treatment has no effect on the expression of TLR4 mRNA in cells induced with LPS.
    Figure Legend Snippet: TLR4 expression in hPVECs and VK2/E6E7 cells. (A). RT-PCR analysis of TLR4 expression in VK2/E6E7 cells and hPVECs. expression of TLR4 (182 bp) in VK2/E6E7 cells (a), (c), and hPVECs (b),(d) was observed. 1: Untreated cells (control); 2: DMSO treated cells (control); 3: TLR4 antibody treated cells; 4: LPS treated cells (10 μg/ml for 6 hrs) and 5: Bay 11–7082 treated cells (5μM for 24 hrs) stimulated with LPS (10μg/ml for 6 hrs) and (c), (d) Housekeeping gene Gapdh (238 bp) considered as an internal standard. The gel picture shown is one of the representative pictures from three independent experiments. (B). Densitometric analysis of bands from RT–PCR amplification products of TLR4 mRNA reported in Fig 10A. The expression of TLR4 mRNA was up-regulated in LPS-induced cells. Bay 11–7082 treatment has no effect on the expression of TLR4 mRNA in cells induced with LPS.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. (A) RT-PCR analysis; 1: Untreated hPVECs, 2: hPVECs treated for 6 hrs with LPS (10 μg/ml), 3: Untreated VK2/E6E7 cells and 4: VK2 cells treated with LPS (10 μg/ml) for 6 hrs. Loading control, Gapdh (238 bp) expression in hPVECs. The gels shown are one of the representative pictures from three independent experiments performed on three different days. (B) Densitometric analysis of bands from RT-PCR amplification products of Hb-α and Hb-β mRNAs shown in figure-5A.
    Figure Legend Snippet: Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. (A) RT-PCR analysis; 1: Untreated hPVECs, 2: hPVECs treated for 6 hrs with LPS (10 μg/ml), 3: Untreated VK2/E6E7 cells and 4: VK2 cells treated with LPS (10 μg/ml) for 6 hrs. Loading control, Gapdh (238 bp) expression in hPVECs. The gels shown are one of the representative pictures from three independent experiments performed on three different days. (B) Densitometric analysis of bands from RT-PCR amplification products of Hb-α and Hb-β mRNAs shown in figure-5A.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    Immunofluorescence localization of vimentin. hPVECs (a-c) and VK2/E6E7 cells (d-f) did not express vimentin. Cytoplasmic localization of vimentin (green) is seen in HeLa cells (positive control) (g-i). Nucleus was stained with DAPI (blue), FITC (a, d, g), FITC and DAPI merge (b,e,h) and no primary antibody controls (c,f,i) are indicated. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).
    Figure Legend Snippet: Immunofluorescence localization of vimentin. hPVECs (a-c) and VK2/E6E7 cells (d-f) did not express vimentin. Cytoplasmic localization of vimentin (green) is seen in HeLa cells (positive control) (g-i). Nucleus was stained with DAPI (blue), FITC (a, d, g), FITC and DAPI merge (b,e,h) and no primary antibody controls (c,f,i) are indicated. The figure shown is one of the representative pictures from three independent experiments (Mag. 63X).

    Techniques Used: Immunofluorescence, Positive Control, Staining

    Expression of NF- κ B in hPVECs and VK2/E6E7 cells. (A). NF-κB levels in un-stimulated, LPS stimulated and Bay 11–7082 treated hPVECs and VK2/E6E7 cells analyzed by ELISA. Cells were seeded at a density of 10 6 /well in 24-well plates and induced with LPS (10 μg/ml for 6 hrs) or Bay-11-0782 (5 μM for 24 hrs). Levels of p65-NF-κB were up-regulated in LPS-induced cells, where as Bay-11-7082 reversed this effect (a). Values represent mean ± SD of three experiments performed on different days. Values are statistically significant (***p
    Figure Legend Snippet: Expression of NF- κ B in hPVECs and VK2/E6E7 cells. (A). NF-κB levels in un-stimulated, LPS stimulated and Bay 11–7082 treated hPVECs and VK2/E6E7 cells analyzed by ELISA. Cells were seeded at a density of 10 6 /well in 24-well plates and induced with LPS (10 μg/ml for 6 hrs) or Bay-11-0782 (5 μM for 24 hrs). Levels of p65-NF-κB were up-regulated in LPS-induced cells, where as Bay-11-7082 reversed this effect (a). Values represent mean ± SD of three experiments performed on different days. Values are statistically significant (***p

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay

    RT-PCR analysis of Hb-α and Hb-β in presence of LPS and Bay 11–7082. (A) Hb-α (429 bp) (a) and Hb-β (444 bp) (b) expression was observed in hPVECs (1,2) and VK2/E6E7 cells (3,4) before and after the treatment with LPS (10 μg/ml for 6 hrs) and Bay 11–7082 (5μM for 24 hrs). LPS-induced (a1, a3, b1, b3), Bay 11–7082 treated (a2, a4, b2, b4) are shown. Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells, whereas Bay 11–7082 attenuated this expression (c) Gapdh (238 bp) was used as loading control. The gels shown are the representative pictures from three independent experiments. (B). Densitometric analysis of RT-PCR bands shown in Fig 12A. Expression of both Hb-α and Hb-β was elevated in LPS-stimulated cells. In contrast, Bay 11–7082 attenuated the expression of Hb-α and Hb-β .
    Figure Legend Snippet: RT-PCR analysis of Hb-α and Hb-β in presence of LPS and Bay 11–7082. (A) Hb-α (429 bp) (a) and Hb-β (444 bp) (b) expression was observed in hPVECs (1,2) and VK2/E6E7 cells (3,4) before and after the treatment with LPS (10 μg/ml for 6 hrs) and Bay 11–7082 (5μM for 24 hrs). LPS-induced (a1, a3, b1, b3), Bay 11–7082 treated (a2, a4, b2, b4) are shown. Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells, whereas Bay 11–7082 attenuated this expression (c) Gapdh (238 bp) was used as loading control. The gels shown are the representative pictures from three independent experiments. (B). Densitometric analysis of RT-PCR bands shown in Fig 12A. Expression of both Hb-α and Hb-β was elevated in LPS-stimulated cells. In contrast, Bay 11–7082 attenuated the expression of Hb-α and Hb-β .

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    Western blot analysis of biomarker expression in hPVECs, VK2/E6E7 cells and RBCs. (a) Cytokeratin-13 (47kDa) expression: Lane-1: RBCs, lane-2: hPVECs and lane-3: VK2/E6E7 cells. (b) Solute carrier family 4 member-1 (SLC4A1) (100 kDa) expression: Lane-1: RBCs (positive control), lane-2: hPVECs, lane-3: VK2/E6E7 cells. (c)Vimentin (54 kDa) expression: Lane-1: vimentin expression in HeLa cells (positive control), Lane-2: hPVECs, Lane-3: VK2/E6E7 cells. (d) β-actin (42 kDa) expression in hPVECs (loading control). The blots shown are one of the representative pictures from three independent experiments performed on three different days.
    Figure Legend Snippet: Western blot analysis of biomarker expression in hPVECs, VK2/E6E7 cells and RBCs. (a) Cytokeratin-13 (47kDa) expression: Lane-1: RBCs, lane-2: hPVECs and lane-3: VK2/E6E7 cells. (b) Solute carrier family 4 member-1 (SLC4A1) (100 kDa) expression: Lane-1: RBCs (positive control), lane-2: hPVECs, lane-3: VK2/E6E7 cells. (c)Vimentin (54 kDa) expression: Lane-1: vimentin expression in HeLa cells (positive control), Lane-2: hPVECs, Lane-3: VK2/E6E7 cells. (d) β-actin (42 kDa) expression in hPVECs (loading control). The blots shown are one of the representative pictures from three independent experiments performed on three different days.

    Techniques Used: Western Blot, Biomarker Assay, Expressing, Positive Control

    RT-PCR analysis of human-β defensin-1 ( hBD-1 ). (A) mRNA expression of hBD1 (139bp) was observed in un-stimulated (a1, a3), LPS stimulated (a2, a4) in hPVECs (a1, a2) and VK2/E6E7 cells (a3, a4). Cells were seeded at a density of 10 6 /well in a 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Expression of hBD1 mRNA was up-regulated in LPS-induced cells. Representative image of RT-PCR analysis of hBD-1 mRNAs expression is shown. Gapdh confirmed roughly equivalent loading of RNA samples. 1: hPVECs (Unstimulated); 2:hPVECs induced with LPS; 3:VK2/E6E7 cells (Unstimulated); 4:VK2/E6E7 cells stimulated with LPS (10μg/ml). (B). Densitometric analysis of bands from RT-PCR amplification products of hBD1 mRNA reported in Figure 13A. Expression of hBD-1 was increased in LPS-stimulated hPVECS and VK2/E6E7 cells.
    Figure Legend Snippet: RT-PCR analysis of human-β defensin-1 ( hBD-1 ). (A) mRNA expression of hBD1 (139bp) was observed in un-stimulated (a1, a3), LPS stimulated (a2, a4) in hPVECs (a1, a2) and VK2/E6E7 cells (a3, a4). Cells were seeded at a density of 10 6 /well in a 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Expression of hBD1 mRNA was up-regulated in LPS-induced cells. Representative image of RT-PCR analysis of hBD-1 mRNAs expression is shown. Gapdh confirmed roughly equivalent loading of RNA samples. 1: hPVECs (Unstimulated); 2:hPVECs induced with LPS; 3:VK2/E6E7 cells (Unstimulated); 4:VK2/E6E7 cells stimulated with LPS (10μg/ml). (B). Densitometric analysis of bands from RT-PCR amplification products of hBD1 mRNA reported in Figure 13A. Expression of hBD-1 was increased in LPS-stimulated hPVECS and VK2/E6E7 cells.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Amplification

    Cytokoine ELISA of hPVECs and VK2/E6E7 cells. IL-6 (A) and IL-10 (B) in the culture supernatants of un-stimulated and LPS stimulated hPVECs and VK2/E6E7 cells at indicated time points (6, 12, 24 hrs). Cells were seeded at a density of 10 6 /well in 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Levels of IL-6 were up-regulated in LPS-induced cells. Values represent the mean ± SD of three independent experiments performed on different days. Level of significance (*p
    Figure Legend Snippet: Cytokoine ELISA of hPVECs and VK2/E6E7 cells. IL-6 (A) and IL-10 (B) in the culture supernatants of un-stimulated and LPS stimulated hPVECs and VK2/E6E7 cells at indicated time points (6, 12, 24 hrs). Cells were seeded at a density of 10 6 /well in 24-well plates and treated with LPS (10 μg/ml for 6 hrs). Levels of IL-6 were up-regulated in LPS-induced cells. Values represent the mean ± SD of three independent experiments performed on different days. Level of significance (*p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells at protein level. (A, B). Immunofluorescence localization of Hb-α and Hb-β. Cytoplasmic localization (green) of Hb-α (A) and Hb-β (B) was observed in hPVECs (a,b,c) and VK2/E6E7 cells (d,e,f). DAPI stained nucleus (blue). a, d: Untreated cells; b,e: LPS treated cells; c,f: Primary antibody controls did not show Hb-α and Hb-β localization. Expression was significantly up-regulated when hPVECs and VK2/E6E7 cells stimulated with LPS (10 μg/ml for 6 hrs) (b, e). The figure shown is one of the representative pictures from three independent experiments performed on three different days (Mag. 63X). (C). Western blot analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. β-actin (42 kDa) (loading control) expression in hPVECs (1: Untreated hPVECs, 2: hPVECs induced with LPS, 3: Untreated VK2/E6E7 cells and 4: VK2/E6E7 cells induced with LPS). (D). Densitometric analysis of bands from western blot of Hb-α and Hb-β shown in Fig 7C.
    Figure Legend Snippet: Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells at protein level. (A, B). Immunofluorescence localization of Hb-α and Hb-β. Cytoplasmic localization (green) of Hb-α (A) and Hb-β (B) was observed in hPVECs (a,b,c) and VK2/E6E7 cells (d,e,f). DAPI stained nucleus (blue). a, d: Untreated cells; b,e: LPS treated cells; c,f: Primary antibody controls did not show Hb-α and Hb-β localization. Expression was significantly up-regulated when hPVECs and VK2/E6E7 cells stimulated with LPS (10 μg/ml for 6 hrs) (b, e). The figure shown is one of the representative pictures from three independent experiments performed on three different days (Mag. 63X). (C). Western blot analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells. β-actin (42 kDa) (loading control) expression in hPVECs (1: Untreated hPVECs, 2: hPVECs induced with LPS, 3: Untreated VK2/E6E7 cells and 4: VK2/E6E7 cells induced with LPS). (D). Densitometric analysis of bands from western blot of Hb-α and Hb-β shown in Fig 7C.

    Techniques Used: Expressing, Immunofluorescence, Staining, Western Blot

    qPCR analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells before and after induction with LPS. Cells seeded at a density of 10 6 /well in 24-well plates were treated with LPS (10 μg/ml for 6 hrs). Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells. Each bar represents the mean ± SD of three independent replicates (***p
    Figure Legend Snippet: qPCR analysis of Hb-α and Hb-β expression in hPVECs and VK2/E6E7 cells before and after induction with LPS. Cells seeded at a density of 10 6 /well in 24-well plates were treated with LPS (10 μg/ml for 6 hrs). Expression of Hb-α and Hb-β was up-regulated in LPS-induced cells. Each bar represents the mean ± SD of three independent replicates (***p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

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    Expressing:

    Article Title: Expression of hemoglobin-α and β subunits in human vaginal epithelial cells and their functional significance
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