human lung adenocarcinoma cell line h1299  (ATCC)


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    ATCC human lung adenocarcinoma cell line h1299
    a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for <t>H1299</t> cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.
    Human Lung Adenocarcinoma Cell Line H1299, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Cathepsin B regulates ovarian reserve quality and quantity via mitophagy by modulating IGF1R turnover"

    Article Title: Cathepsin B regulates ovarian reserve quality and quantity via mitophagy by modulating IGF1R turnover

    Journal: bioRxiv

    doi: 10.1101/2024.02.14.580410

    a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for H1299 cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.
    Figure Legend Snippet: a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for H1299 cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.

    Techniques Used: Western Blot, In Vitro, Mass Spectrometry, Cell Culture

    human lung adenocarcinoma cell line  (ATCC)


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    ATCC human lung adenocarcinoma cell line
    Human Lung Adenocarcinoma Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human lung adenocarcinoma cell line a549  (ATCC)


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    ATCC human lung adenocarcinoma cell line a549
    Viability of <t>A549</t> cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).
    Human Lung Adenocarcinoma Cell Line A549, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    human lung adenocarcinoma cell line a549 - by Bioz Stars, 2024-02
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    1) Product Images from "Chitosan and Sodium Hyaluronate Hydrogels Supplemented with Bioglass for Bone Tissue Engineering"

    Article Title: Chitosan and Sodium Hyaluronate Hydrogels Supplemented with Bioglass for Bone Tissue Engineering

    Journal: Gels

    doi: 10.3390/gels10020128

    Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).
    Figure Legend Snippet: Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).

    Techniques Used: Cell Culture

    Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 10% wt. of sodium hyaluronate, 45% wt. of chitosan, and 45% wt. of bioglass treated at 550 °C/3 h (C–H10–P5, C–H10–P5Zn2, C–H10–P5Sr2) or bioglass treated at 650 °C/10 h (C–H10–P5d, C–H10–P5Sr2d).
    Figure Legend Snippet: Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 10% wt. of sodium hyaluronate, 45% wt. of chitosan, and 45% wt. of bioglass treated at 550 °C/3 h (C–H10–P5, C–H10–P5Zn2, C–H10–P5Sr2) or bioglass treated at 650 °C/10 h (C–H10–P5d, C–H10–P5Sr2d).

    Techniques Used: Cell Culture

    human lung adenocarcinoma cell line a549  (ATCC)


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    ATCC human lung adenocarcinoma cell line a549
    M. orbicularis’ plant parts’ ethanolic extracts reduce the viability of <t>A549</t> cells. MTT assay was used to measure cell viability of A549 cells treated for 24, 48, and 72 h with increasing concentrations (50, 100, 150, 200 μg/mL) of M. orbicularis’ plant parts’ ethanolic extracts. ( A – C ) show the results of MTT cell cytotoxicity assay of A549 cells treated with the indicated concentrations of ethanolic plant extracts of M. orbicularis leaves, fruits, and stems, respectively. ( D ) shows the cell viability of neonatal fibroblast cells treated with the indicated concentrations of M. orbicularis’ fruits’ ethanolic extract. Viability of treated cells was compared to the control-(ctr) vehicle-treated cells. Data are displayed as the mean ± SEM of three independent experiments (n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001.
    Human Lung Adenocarcinoma Cell Line A549, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human lung adenocarcinoma cell line a549 - by Bioz Stars, 2024-02
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    1) Product Images from "Determination of Medicago orbicularis Antioxidant, Antihemolytic, and Anti-Cancerous Activities and Its Augmentation of Cisplatin-Induced Cytotoxicity in A549 Lung Cancer Cells"

    Article Title: Determination of Medicago orbicularis Antioxidant, Antihemolytic, and Anti-Cancerous Activities and Its Augmentation of Cisplatin-Induced Cytotoxicity in A549 Lung Cancer Cells

    Journal: Plants

    doi: 10.3390/plants13030442

    M. orbicularis’ plant parts’ ethanolic extracts reduce the viability of A549 cells. MTT assay was used to measure cell viability of A549 cells treated for 24, 48, and 72 h with increasing concentrations (50, 100, 150, 200 μg/mL) of M. orbicularis’ plant parts’ ethanolic extracts. ( A – C ) show the results of MTT cell cytotoxicity assay of A549 cells treated with the indicated concentrations of ethanolic plant extracts of M. orbicularis leaves, fruits, and stems, respectively. ( D ) shows the cell viability of neonatal fibroblast cells treated with the indicated concentrations of M. orbicularis’ fruits’ ethanolic extract. Viability of treated cells was compared to the control-(ctr) vehicle-treated cells. Data are displayed as the mean ± SEM of three independent experiments (n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001.
    Figure Legend Snippet: M. orbicularis’ plant parts’ ethanolic extracts reduce the viability of A549 cells. MTT assay was used to measure cell viability of A549 cells treated for 24, 48, and 72 h with increasing concentrations (50, 100, 150, 200 μg/mL) of M. orbicularis’ plant parts’ ethanolic extracts. ( A – C ) show the results of MTT cell cytotoxicity assay of A549 cells treated with the indicated concentrations of ethanolic plant extracts of M. orbicularis leaves, fruits, and stems, respectively. ( D ) shows the cell viability of neonatal fibroblast cells treated with the indicated concentrations of M. orbicularis’ fruits’ ethanolic extract. Viability of treated cells was compared to the control-(ctr) vehicle-treated cells. Data are displayed as the mean ± SEM of three independent experiments (n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001.

    Techniques Used: MTT Assay, Cytotoxicity Assay

    IC50 values (in μg/mL) of ethanolic extracts of parts of M. orbicularis used to treat  A549  cells for 24, 48, and 72 h. The IC50 values of cisplatin in  A549  cells are also listed for comparison.
    Figure Legend Snippet: IC50 values (in μg/mL) of ethanolic extracts of parts of M. orbicularis used to treat A549 cells for 24, 48, and 72 h. The IC50 values of cisplatin in A549 cells are also listed for comparison.

    Techniques Used: Comparison

    M. orbicularis’ fruits’ ethanolic extracts induce apoptosis of A549 cells. ( A ) Representative Western blot of protein lysates from A549 cells treated 100 and 150 μg/mL of M. orbicularis fruit extracts for 48 h. Levels of proteins were normalized to GAPDH protein levels. ( B ) Quantification of images in ( A ). Intensity of protein bands were quantified by ImageJ software and normalized to intensity of bands of GAPDH protein. The ratio is expressed in arbitrary units. Data are presented as the mean ± SEM of 3 independent experiments (n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001.
    Figure Legend Snippet: M. orbicularis’ fruits’ ethanolic extracts induce apoptosis of A549 cells. ( A ) Representative Western blot of protein lysates from A549 cells treated 100 and 150 μg/mL of M. orbicularis fruit extracts for 48 h. Levels of proteins were normalized to GAPDH protein levels. ( B ) Quantification of images in ( A ). Intensity of protein bands were quantified by ImageJ software and normalized to intensity of bands of GAPDH protein. The ratio is expressed in arbitrary units. Data are presented as the mean ± SEM of 3 independent experiments (n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001.

    Techniques Used: Western Blot, Software

    M. orbicularis enhance cisplatin-induced cytotoxicity in A549 lung cancer cells. ( A ) Cell viability of A549 cells treated with the indicated concentrations of cisplatin was measured using MTT assay. Data are displayed as % cell viability relative to control (ctr) cells. * p < 0.05, *** p < 0.001, and **** p < 0.0001 ( B ) A549 cells were treated with fruit extracts of M. orbicularis (100 μg/mL) alone or combined with 5 μg/mL cisplatin (Cis) for 48 h, and then assayed for cell viability using MTT assay. Bar graphs represent % cell viability relative to control (ctr) cells. Data are presented as the mean ± SEM of three independent experiments (n = 3). Significant difference from control: **** p < 0.0001; significant difference from cisplatin treatment alone: #### denotes p < 0.0001; and significant difference from M. orbicularis alone: ‡ denotes p < 0.05. M. orbicularis F. denotes M. orbicularis fruits extracts.
    Figure Legend Snippet: M. orbicularis enhance cisplatin-induced cytotoxicity in A549 lung cancer cells. ( A ) Cell viability of A549 cells treated with the indicated concentrations of cisplatin was measured using MTT assay. Data are displayed as % cell viability relative to control (ctr) cells. * p < 0.05, *** p < 0.001, and **** p < 0.0001 ( B ) A549 cells were treated with fruit extracts of M. orbicularis (100 μg/mL) alone or combined with 5 μg/mL cisplatin (Cis) for 48 h, and then assayed for cell viability using MTT assay. Bar graphs represent % cell viability relative to control (ctr) cells. Data are presented as the mean ± SEM of three independent experiments (n = 3). Significant difference from control: **** p < 0.0001; significant difference from cisplatin treatment alone: #### denotes p < 0.0001; and significant difference from M. orbicularis alone: ‡ denotes p < 0.05. M. orbicularis F. denotes M. orbicularis fruits extracts.

    Techniques Used: MTT Assay

    M. orbicularis augments cisplatin-induced aggregation of A549 lung cancer cells. ( A ) A549 cells were incubated with M. orbicularis alone (100 or 150 μg/mL) or combined with 5 μg/mL cisplatin (Cis), and then subjected to cell aggregation assay as described in the Materials and Methods Section. ( B ) Quantification of the data in ( A ). Micrographs of cells were taken after 1 h of treatment and the percentage of cell–cell aggregations was measured using the following equation: % aggregation = (1 − Nt/Nc) × 100, where Nt is the number of single cells in the control and Nc is the number of single cells in the treated sample. Data represent the mean ± SEM of three independent experiments (n = 3). Significant difference from control: *** p < 0.001, and **** p < 0.0001; significant difference from cisplatin treatment alone: # denotes p < 0.005; and significant difference from M. orbicularis extracts alone: ‡ denotes p < 0.05. M. orbicularis F. denotes M. orbicularis’ fruits’ extracts.
    Figure Legend Snippet: M. orbicularis augments cisplatin-induced aggregation of A549 lung cancer cells. ( A ) A549 cells were incubated with M. orbicularis alone (100 or 150 μg/mL) or combined with 5 μg/mL cisplatin (Cis), and then subjected to cell aggregation assay as described in the Materials and Methods Section. ( B ) Quantification of the data in ( A ). Micrographs of cells were taken after 1 h of treatment and the percentage of cell–cell aggregations was measured using the following equation: % aggregation = (1 − Nt/Nc) × 100, where Nt is the number of single cells in the control and Nc is the number of single cells in the treated sample. Data represent the mean ± SEM of three independent experiments (n = 3). Significant difference from control: *** p < 0.001, and **** p < 0.0001; significant difference from cisplatin treatment alone: # denotes p < 0.005; and significant difference from M. orbicularis extracts alone: ‡ denotes p < 0.05. M. orbicularis F. denotes M. orbicularis’ fruits’ extracts.

    Techniques Used: Incubation

    human lung adenocarcinoma cell line  (ATCC)


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    ATCC human lung adenocarcinoma cell line
    Human Lung Adenocarcinoma Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human lung adenocarcinoma a549 cell line  (ATCC)


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    ATCC human lung adenocarcinoma a549 cell line
    The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of <t>A549</t> cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.
    Human Lung Adenocarcinoma A549 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human lung adenocarcinoma a549 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
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    1) Product Images from "Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway"

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    Journal: Journal of Thoracic Disease

    doi: 10.21037/jtd-23-1171

    The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of A549 cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.
    Figure Legend Snippet: The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of A549 cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.

    Techniques Used: CCK-8 Assay, Expressing, Concentration Assay, Cell Counting, Real-time Polymerase Chain Reaction

    The expression levels of EGR4 and OCT4 in chemo-resistant cell lines and tissues from patients before and after DDP-chemotherapy (*, P<0.05). (A,B) Real-time PCR confirmed that markedly up-regulation of EGR4 and OCT4, respectively, in A549/DDP cells, as opposed to that in A549 cells and BEAS-2B cells. (C,D) IHC results found that protein expression levels of EGR4 and OCT4, respectively, were notably higher in DDP-resistant tissues (the scale bars in IHC represent 50 μm) (left: post-DDP therapy; right: pre-DDP therapy). DDP, cisplatin; PCR, polymerase chain reaction; IHC, immunohistochemistry.
    Figure Legend Snippet: The expression levels of EGR4 and OCT4 in chemo-resistant cell lines and tissues from patients before and after DDP-chemotherapy (*, P<0.05). (A,B) Real-time PCR confirmed that markedly up-regulation of EGR4 and OCT4, respectively, in A549/DDP cells, as opposed to that in A549 cells and BEAS-2B cells. (C,D) IHC results found that protein expression levels of EGR4 and OCT4, respectively, were notably higher in DDP-resistant tissues (the scale bars in IHC represent 50 μm) (left: post-DDP therapy; right: pre-DDP therapy). DDP, cisplatin; PCR, polymerase chain reaction; IHC, immunohistochemistry.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Immunohistochemistry

    Knockdown of ZNF205-AS1 led to elevated miRNA-138-5p levels in A549/DDP cells. (A) The down-regulation of ZNF205-AS1 was realized by transfecting three designed small interfering RNAs. ZNF205-AS1 level was significantly higher in blank control group and NC group than the three paralleled knockdown group (***, P<0.001). (B) The levels of miR-138-5p in A549/DDP cells with ZNF205-AS1 loss-of-function were assessed in qPCR. ZNF205-AS1 loss-of-function led to significantly up-regulation of miR-138-5p as opposed to NC group (**, P<0.01) (GAPDH and U6 were used as endogenous control for detection of ZNF205-AS1 and miRNA-138-5p, respectively). (C) The binding site of lncRNA ZNF205-AS1 with miRNA-138-5p was predicted in bioinformatics analysis. (D) The significantly inhibited relative luciferase activity of cells co-transfected with wild-type ZNF205-AS1 and miRNA-138-5p mimic, as opposed to that transfected with wild-type ZNF205-AS1 and miR-NC was detected (**, P<0.01). NC, negative control; WT, wide type; Mut, mutant; DDP, cisplatin; qPCR, quantitative polymerase chain reaction.
    Figure Legend Snippet: Knockdown of ZNF205-AS1 led to elevated miRNA-138-5p levels in A549/DDP cells. (A) The down-regulation of ZNF205-AS1 was realized by transfecting three designed small interfering RNAs. ZNF205-AS1 level was significantly higher in blank control group and NC group than the three paralleled knockdown group (***, P<0.001). (B) The levels of miR-138-5p in A549/DDP cells with ZNF205-AS1 loss-of-function were assessed in qPCR. ZNF205-AS1 loss-of-function led to significantly up-regulation of miR-138-5p as opposed to NC group (**, P<0.01) (GAPDH and U6 were used as endogenous control for detection of ZNF205-AS1 and miRNA-138-5p, respectively). (C) The binding site of lncRNA ZNF205-AS1 with miRNA-138-5p was predicted in bioinformatics analysis. (D) The significantly inhibited relative luciferase activity of cells co-transfected with wild-type ZNF205-AS1 and miRNA-138-5p mimic, as opposed to that transfected with wild-type ZNF205-AS1 and miR-NC was detected (**, P<0.01). NC, negative control; WT, wide type; Mut, mutant; DDP, cisplatin; qPCR, quantitative polymerase chain reaction.

    Techniques Used: Binding Assay, Luciferase, Activity Assay, Transfection, Negative Control, Mutagenesis, Real-time Polymerase Chain Reaction

    miRNA-138-5p gain-of-function enhanced the sensitivity of A549/DDP cells to DDP. (A) Overexpression of miRNA-138-5p was verified in qPCR assay to find that miRNA-138-5p level was significantly higher in miRNA-138-5p mimic group than NC group (*, P<0.05) (U6 as endogenous control). (B) Up-regulation of miR-138-5p promoted the invasion of A549/DDP cells. The cells were stained with 0.1% Giemsa stain (the scale bars represent 25 μm). (C) The results of CCK8 assay showed that the viability of cells exposed to gradient concentrations of DDP was significantly inhibited in miRNA-138-5p mimic group, compared with that of NC group (*, P<0.05). DDP, cisplatin; NC, negative control; IC 50 , half-maximal inhibitory concentration; qPCR, quantitative polymerase chain reaction; CCK8, Cell Counting Kit-8.
    Figure Legend Snippet: miRNA-138-5p gain-of-function enhanced the sensitivity of A549/DDP cells to DDP. (A) Overexpression of miRNA-138-5p was verified in qPCR assay to find that miRNA-138-5p level was significantly higher in miRNA-138-5p mimic group than NC group (*, P<0.05) (U6 as endogenous control). (B) Up-regulation of miR-138-5p promoted the invasion of A549/DDP cells. The cells were stained with 0.1% Giemsa stain (the scale bars represent 25 μm). (C) The results of CCK8 assay showed that the viability of cells exposed to gradient concentrations of DDP was significantly inhibited in miRNA-138-5p mimic group, compared with that of NC group (*, P<0.05). DDP, cisplatin; NC, negative control; IC 50 , half-maximal inhibitory concentration; qPCR, quantitative polymerase chain reaction; CCK8, Cell Counting Kit-8.

    Techniques Used: Over Expression, Staining, Giemsa Stain, CCK-8 Assay, Negative Control, Concentration Assay, Real-time Polymerase Chain Reaction, Cell Counting

    miR-138-5p up-regulation facilitated DDP-induced apoptosis of A549/DDP cells. (A) The cell apoptosis rate of DDP-treated A549/DDP cells with miR-138-5p overexpression was significantly increased than those without miR-138-5p overexpression (*, P<0.05). (B) The protein levels of Cleaved-caspase 3, Cleaved-PARP, Bax, Bcl2, and OCT4 were measured in western blotting (*, P<0.05). (C) RNA pulldown assay was performed to verify the potential binding of miR-138-5p to OCT4, and EGR4, respectively (lane 1: miR-NC input; lane 2: miR-138-input; lane 3: miR-NC pulldown; lane 4: miR-138-5p pulldown). NC, negative control; OCT4, octamer-binding protein 4; EGR4, early growth response 4; DDP, cisplatin.
    Figure Legend Snippet: miR-138-5p up-regulation facilitated DDP-induced apoptosis of A549/DDP cells. (A) The cell apoptosis rate of DDP-treated A549/DDP cells with miR-138-5p overexpression was significantly increased than those without miR-138-5p overexpression (*, P<0.05). (B) The protein levels of Cleaved-caspase 3, Cleaved-PARP, Bax, Bcl2, and OCT4 were measured in western blotting (*, P<0.05). (C) RNA pulldown assay was performed to verify the potential binding of miR-138-5p to OCT4, and EGR4, respectively (lane 1: miR-NC input; lane 2: miR-138-input; lane 3: miR-NC pulldown; lane 4: miR-138-5p pulldown). NC, negative control; OCT4, octamer-binding protein 4; EGR4, early growth response 4; DDP, cisplatin.

    Techniques Used: Over Expression, Western Blot, Binding Assay, Negative Control

    miRNA-138-5p loss-of-function diminished the pro-apoptosis activity induced by ZNF205-AS1-silencing in A549/DDP cells. (A) CCK8 assay confirmed that the cell viability of si-ZNF205-AS1 group was significantly lower than that with miR-138-5p knockdown and NC group (*, P<0.05). (B) Significantly higher protein levels of OCT4 and Bcl-2, as well as significantly lower protein levels of Cl-caspase-3, Cl-PARP and Bax were detected in si-ZNF205-AS1 with loss-of-function of miR-138-5p than that without (*, P<0.05). (C) The apoptosis rate of si-ZNF205-AS1 group was significantly higher than that of si-ZNF205-AS1 + miRNA-138-5p group and NC group (*, P<0.05). (D) The number of invasive cells was significantly higher in si-ZNF205-AS1 + miRNA-138-5p group and in NC group, in comparison with si-ZNF205-AS1 group (*, P<0.05). The cells were stained with 0.1% Giemsa stain (magnification 200×). (E) The migration ability of si-ZNF205-AS1 group being significantly lower than si-ZNF205-AS1 + miRNA-138-5p group (**, P<0.01) and NC group was validated in wound healing assay (the scratched areas from each group at 0 and 24 h post-wound were photographed and quantified on a microscope) (***, P<0.001). NC, negative control; DDP, cisplatin; CCK8, Cell Counting Kit-8.
    Figure Legend Snippet: miRNA-138-5p loss-of-function diminished the pro-apoptosis activity induced by ZNF205-AS1-silencing in A549/DDP cells. (A) CCK8 assay confirmed that the cell viability of si-ZNF205-AS1 group was significantly lower than that with miR-138-5p knockdown and NC group (*, P<0.05). (B) Significantly higher protein levels of OCT4 and Bcl-2, as well as significantly lower protein levels of Cl-caspase-3, Cl-PARP and Bax were detected in si-ZNF205-AS1 with loss-of-function of miR-138-5p than that without (*, P<0.05). (C) The apoptosis rate of si-ZNF205-AS1 group was significantly higher than that of si-ZNF205-AS1 + miRNA-138-5p group and NC group (*, P<0.05). (D) The number of invasive cells was significantly higher in si-ZNF205-AS1 + miRNA-138-5p group and in NC group, in comparison with si-ZNF205-AS1 group (*, P<0.05). The cells were stained with 0.1% Giemsa stain (magnification 200×). (E) The migration ability of si-ZNF205-AS1 group being significantly lower than si-ZNF205-AS1 + miRNA-138-5p group (**, P<0.01) and NC group was validated in wound healing assay (the scratched areas from each group at 0 and 24 h post-wound were photographed and quantified on a microscope) (***, P<0.001). NC, negative control; DDP, cisplatin; CCK8, Cell Counting Kit-8.

    Techniques Used: Activity Assay, CCK-8 Assay, Comparison, Staining, Giemsa Stain, Migration, Wound Healing Assay, Microscopy, Negative Control, Cell Counting

    human lung adenocarcinoma cell line a549  (ATCC)


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    ATCC human lung adenocarcinoma cell line a549
    Human Lung Adenocarcinoma Cell Line A549, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human lung adenocarcinoma cell line calu 3  (ATCC)


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    ATCC human lung adenocarcinoma cell line calu 3
    (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial <t>CaLu-3</t> cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.
    Human Lung Adenocarcinoma Cell Line Calu 3, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants"

    Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

    Journal: Cell

    doi: 10.1016/j.cell.2023.12.026

    (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial CaLu-3 cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.
    Figure Legend Snippet: (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial CaLu-3 cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

    Techniques Used: Binding Assay, Membrane, Infection, Stable Transfection, Expressing, Derivative Assay, Transfection

    (A–D) HEK293T cells were cotransfected with the indicated spikes of interest and GFP plasmids and were cocultured with 293T-ACE2 (A-B) or human lung epithelial CaLu-3 cells (C-D) for 24 h. Cell-cell fusion was imaged and GFP areas of fused cells were quantified (see ). D614G and no S were included as positive and negative control, respectively. Comparisons in extents of cell-cell fusion for each Omicron subvariant were made against D614G. Scale bars represent 150 μM. Bars in (B and D) represent means ± standard error. Dots represent three images from two biological replicates. Statistical significance relative to D614G was determined using a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 3). p values are displayed as ns p > 0.05, *p < 0.05, ***p < 0.001, and ****p < 0.0001.
    Figure Legend Snippet: (A–D) HEK293T cells were cotransfected with the indicated spikes of interest and GFP plasmids and were cocultured with 293T-ACE2 (A-B) or human lung epithelial CaLu-3 cells (C-D) for 24 h. Cell-cell fusion was imaged and GFP areas of fused cells were quantified (see ). D614G and no S were included as positive and negative control, respectively. Comparisons in extents of cell-cell fusion for each Omicron subvariant were made against D614G. Scale bars represent 150 μM. Bars in (B and D) represent means ± standard error. Dots represent three images from two biological replicates. Statistical significance relative to D614G was determined using a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 3). p values are displayed as ns p > 0.05, *p < 0.05, ***p < 0.001, and ****p < 0.0001.

    Techniques Used: Negative Control

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Infection, Recombinant, Transfection, Modification, Protease Inhibitor, Western Blot, Software, Imaging

    human lung adenocarcinoma cell line calu 3  (ATCC)


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    ATCC human lung adenocarcinoma cell line calu 3
    Human Lung Adenocarcinoma Cell Line Calu 3, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human lung adenocarcinoma a549 cell line  (ATCC)


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    ATCC human lung adenocarcinoma a549 cell line
    Fraisinib treatment induces caspase cascade activation, cell cycle arrests at the G0/G1 phase, and apoptosis activation and invasiveness inhibition in <t>A549</t> cells. (A) FACS-based activation assays of the multicaspase complex in A549 cells. Profiles were determined in untreated cells (control) and after 48 h treatment with DMSO or 10 μM Fraisinib. (B) Bar charts depict the percentage of live cells and multicaspase enzyme activation. (C) Relative fractions of cells in G0/G1, S, and G2/M stages determined for cells exposed to DMSO or 10 μM Fraisinib. Statistical difference between DMSO- and Fraisinib-treated cells in G0/G1 was calculated by paired Student’s t-test (asterisk: p -value = 0.01). (D) Comparison of the expression level of apoptotic proteins extracted from DMSO- and Fraisinib-treated A549 cells. Results are the mean ± SD of two independent wells within the array; asterisks indicate the protein level ratio between treated vs. untreated cells higher than 20%. (E) Bar diagram showing the percentage of damaged cells revealed by c.Live Tox reagent 250 nM (Cytena) in DMSO- and Fraisinib-treated cells after 48 h treatments. Values are the mean ± SD of three experiments performed in quadruplicate. (F) Images representing cell migration in three independent experiments performed to evaluate invasion inhibition in A549 cells with added DMSO (top images) or exposed to Fraisinib (bottom images). The bars correspond to 100 μm. (G) Histogram reporting the mean value ± SD of the migration index of the three independent experiments (asterisks: p -value< 0.001).
    Human Lung Adenocarcinoma A549 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Fraisinib: a calixpyrrole derivative reducing A549 cell-derived NSCLC tumor in vivo acts as a ligand of the glycine-tRNA synthase, a new molecular target in oncology"

    Article Title: Fraisinib: a calixpyrrole derivative reducing A549 cell-derived NSCLC tumor in vivo acts as a ligand of the glycine-tRNA synthase, a new molecular target in oncology

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2023.1258108

    Fraisinib treatment induces caspase cascade activation, cell cycle arrests at the G0/G1 phase, and apoptosis activation and invasiveness inhibition in A549 cells. (A) FACS-based activation assays of the multicaspase complex in A549 cells. Profiles were determined in untreated cells (control) and after 48 h treatment with DMSO or 10 μM Fraisinib. (B) Bar charts depict the percentage of live cells and multicaspase enzyme activation. (C) Relative fractions of cells in G0/G1, S, and G2/M stages determined for cells exposed to DMSO or 10 μM Fraisinib. Statistical difference between DMSO- and Fraisinib-treated cells in G0/G1 was calculated by paired Student’s t-test (asterisk: p -value = 0.01). (D) Comparison of the expression level of apoptotic proteins extracted from DMSO- and Fraisinib-treated A549 cells. Results are the mean ± SD of two independent wells within the array; asterisks indicate the protein level ratio between treated vs. untreated cells higher than 20%. (E) Bar diagram showing the percentage of damaged cells revealed by c.Live Tox reagent 250 nM (Cytena) in DMSO- and Fraisinib-treated cells after 48 h treatments. Values are the mean ± SD of three experiments performed in quadruplicate. (F) Images representing cell migration in three independent experiments performed to evaluate invasion inhibition in A549 cells with added DMSO (top images) or exposed to Fraisinib (bottom images). The bars correspond to 100 μm. (G) Histogram reporting the mean value ± SD of the migration index of the three independent experiments (asterisks: p -value< 0.001).
    Figure Legend Snippet: Fraisinib treatment induces caspase cascade activation, cell cycle arrests at the G0/G1 phase, and apoptosis activation and invasiveness inhibition in A549 cells. (A) FACS-based activation assays of the multicaspase complex in A549 cells. Profiles were determined in untreated cells (control) and after 48 h treatment with DMSO or 10 μM Fraisinib. (B) Bar charts depict the percentage of live cells and multicaspase enzyme activation. (C) Relative fractions of cells in G0/G1, S, and G2/M stages determined for cells exposed to DMSO or 10 μM Fraisinib. Statistical difference between DMSO- and Fraisinib-treated cells in G0/G1 was calculated by paired Student’s t-test (asterisk: p -value = 0.01). (D) Comparison of the expression level of apoptotic proteins extracted from DMSO- and Fraisinib-treated A549 cells. Results are the mean ± SD of two independent wells within the array; asterisks indicate the protein level ratio between treated vs. untreated cells higher than 20%. (E) Bar diagram showing the percentage of damaged cells revealed by c.Live Tox reagent 250 nM (Cytena) in DMSO- and Fraisinib-treated cells after 48 h treatments. Values are the mean ± SD of three experiments performed in quadruplicate. (F) Images representing cell migration in three independent experiments performed to evaluate invasion inhibition in A549 cells with added DMSO (top images) or exposed to Fraisinib (bottom images). The bars correspond to 100 μm. (G) Histogram reporting the mean value ± SD of the migration index of the three independent experiments (asterisks: p -value< 0.001).

    Techniques Used: Activation Assay, Inhibition, Comparison, Expressing, Migration

    GARS1 interactome in the A549 cell line. Network parameters are reported in the inset table.
    Figure Legend Snippet: GARS1 interactome in the A549 cell line. Network parameters are reported in the inset table.

    Techniques Used:

    Timeline of the biological mechanisms dysregulated in the A549 cells following Fraisinib treatment. The chronological progression of pathway dysregulation, observed by comparing the DMSO control and Fraisinib-treated cells at 24, 48, and 72 h, is shown over time. Under the timeline, each strip spans the first and last treatment time point at which the biological mechanism resulted dysregulated; the strips are color coded as follows: red for biological mechanisms altered at all time points; blue at 24 and 48 h, green only at 24 h, and violet at 48 and 72 h. The colors at the edges of each strip are faded as the data were obtained at definite time points and both the start and the end of each process cannot be exactly defined. General biological mechanisms are written in black, and cancer hallmarks are written in bold white; the biological mechanisms specifically regulated by GARS1 are indicated with an asterisk (retrieved from Cancer Gene Net, https://signor.uniroma2.it/CancerGeneNet/ ).
    Figure Legend Snippet: Timeline of the biological mechanisms dysregulated in the A549 cells following Fraisinib treatment. The chronological progression of pathway dysregulation, observed by comparing the DMSO control and Fraisinib-treated cells at 24, 48, and 72 h, is shown over time. Under the timeline, each strip spans the first and last treatment time point at which the biological mechanism resulted dysregulated; the strips are color coded as follows: red for biological mechanisms altered at all time points; blue at 24 and 48 h, green only at 24 h, and violet at 48 and 72 h. The colors at the edges of each strip are faded as the data were obtained at definite time points and both the start and the end of each process cannot be exactly defined. General biological mechanisms are written in black, and cancer hallmarks are written in bold white; the biological mechanisms specifically regulated by GARS1 are indicated with an asterisk (retrieved from Cancer Gene Net, https://signor.uniroma2.it/CancerGeneNet/ ).

    Techniques Used: Stripping Membranes

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    ATCC human lung adenocarcinoma cell line h1299
    a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for <t>H1299</t> cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.
    Human Lung Adenocarcinoma Cell Line H1299, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human lung adenocarcinoma cell line
    a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for <t>H1299</t> cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.
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    ATCC human lung adenocarcinoma cell line a549
    Viability of <t>A549</t> cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).
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    ATCC human lung adenocarcinoma a549 cell line
    The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of <t>A549</t> cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.
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    ATCC human lung adenocarcinoma cell line calu 3
    (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial <t>CaLu-3</t> cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.
    Human Lung Adenocarcinoma Cell Line Calu 3, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for H1299 cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.

    Journal: bioRxiv

    Article Title: Cathepsin B regulates ovarian reserve quality and quantity via mitophagy by modulating IGF1R turnover

    doi: 10.1101/2024.02.14.580410

    Figure Lengend Snippet: a, Venn diagram of VC and i_Cat B -treated ovaries proteome. b , Principal Component Analysis (PCA) shows the difference between the VC and i_Cat B -treated ovaries proteome profiles. c, Relative abundance of differential proteins in VC and i_Cat B -treated ovaries presented as a heatmap. d, Distribution of all protein classes identified in VC and i_Cat B - treated ovaries according to biological process. e, A volcano plot showing P values versus fold changes of all proteins in VC and i_Cat B -treated ovaries. f , Schematic representation of insulin signalling pathway in oocytes. g, Western blot analysis of IGF1, IGF1R, pIGF1R, pAKT, pMTOR, Cat B, MVH and Tubulin from VC and i_Cat B -treated ovaries. h , Experimental regimen for in vitro reaction followed by mass spectrometry. i , Western blot analysis of in vitro digestion by Cat B. j , Schematic representation of invitro digested peptides of IGF1R identified in mass spectrometry. k , Experimental regimen for H1299 cell culture experiments. l, Western blot analysis of Cat B, IGF1R, and Actin from Control and Cat B-overexpressed H1299 cells. m, Western blot analysis of Cat B, IGF1R, and Actin from Control and i_Cat B -treated H1299 cells. VC, vehicle control; i_Cat B, an inhibitor of Cathepsin B.

    Article Snippet: Human lung adenocarcinoma cell line H1299 purchased from ATCC (ATCC - CRL-5803) was grown in RPMI1640 (Gibco - 11875085), 10% FBS (Gibco - 10270106) and 1× penicillin– streptomycin (Sigma -P4333) at 37 °C and 5% CO2.

    Techniques: Western Blot, In Vitro, Mass Spectrometry, Cell Culture

    Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).

    Journal: Gels

    Article Title: Chitosan and Sodium Hyaluronate Hydrogels Supplemented with Bioglass for Bone Tissue Engineering

    doi: 10.3390/gels10020128

    Figure Lengend Snippet: Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 5% wt. of sodium hyaluronate, 47.5% wt. of chitosan, and 47.5% wt. of bioglass treated at 550 °C/3 h (C–H5-P5, C–H5–P5Zn2, C–H5–P5Sr2) or bioglass treated at 650 °C/10 h (C–H5–P5d, C–H5–P5Zn2d, C–H5–P5Sr2d).

    Article Snippet: The human lung adenocarcinoma cell line A549 (ATCC CCL 185) was derived from the Institute of Immunology and Experimental Therapy collection of cell lines (Wrocław, Poland), while MG63 cells (human osteoblast-like MG63 cells) were from the European Collection of Cell Cultures, Salisbury, UK.

    Techniques: Cell Culture

    Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 10% wt. of sodium hyaluronate, 45% wt. of chitosan, and 45% wt. of bioglass treated at 550 °C/3 h (C–H10–P5, C–H10–P5Zn2, C–H10–P5Sr2) or bioglass treated at 650 °C/10 h (C–H10–P5d, C–H10–P5Sr2d).

    Journal: Gels

    Article Title: Chitosan and Sodium Hyaluronate Hydrogels Supplemented with Bioglass for Bone Tissue Engineering

    doi: 10.3390/gels10020128

    Figure Lengend Snippet: Viability of A549 cells (assessed via MTT test) cultured in extracts from composites containing 10% wt. of sodium hyaluronate, 45% wt. of chitosan, and 45% wt. of bioglass treated at 550 °C/3 h (C–H10–P5, C–H10–P5Zn2, C–H10–P5Sr2) or bioglass treated at 650 °C/10 h (C–H10–P5d, C–H10–P5Sr2d).

    Article Snippet: The human lung adenocarcinoma cell line A549 (ATCC CCL 185) was derived from the Institute of Immunology and Experimental Therapy collection of cell lines (Wrocław, Poland), while MG63 cells (human osteoblast-like MG63 cells) were from the European Collection of Cell Cultures, Salisbury, UK.

    Techniques: Cell Culture

    The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of A549 cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: The levels of ZNF205-AS1 and miRNA-138-5p in non-resistant NSCLC cells and DDP-resistant cells (*, P<0.05 and **, P<0.01). (A) The biological effect of various concentrations of DDP on the viability of A549 cells and A549/DDP cells was measured in CCK8 assay, which showed that the IC 50 value of A549 cell line was significantly lower than that of A549/DDP (10.28±0.92 vs. 38.13±5.72 μM; P<0.001). (B) qPCR was conducted to evaluate the expression of ZNF205-AS1 in DDP-sensitive A549 cells and A549/DDP cells (GAPDH as endogenous control). (C) qPCR was employed to evaluate the expression of miRNA-138-5p in DDP sensitive A549 cells and A549/DDP cells (U6 as endogenous control). DDP, cisplatin; IC 50 , half-maximal inhibitory concentration; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit-8; qPCR, quantitative polymerase chain reaction.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: CCK-8 Assay, Expressing, Concentration Assay, Cell Counting, Real-time Polymerase Chain Reaction

    The expression levels of EGR4 and OCT4 in chemo-resistant cell lines and tissues from patients before and after DDP-chemotherapy (*, P<0.05). (A,B) Real-time PCR confirmed that markedly up-regulation of EGR4 and OCT4, respectively, in A549/DDP cells, as opposed to that in A549 cells and BEAS-2B cells. (C,D) IHC results found that protein expression levels of EGR4 and OCT4, respectively, were notably higher in DDP-resistant tissues (the scale bars in IHC represent 50 μm) (left: post-DDP therapy; right: pre-DDP therapy). DDP, cisplatin; PCR, polymerase chain reaction; IHC, immunohistochemistry.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: The expression levels of EGR4 and OCT4 in chemo-resistant cell lines and tissues from patients before and after DDP-chemotherapy (*, P<0.05). (A,B) Real-time PCR confirmed that markedly up-regulation of EGR4 and OCT4, respectively, in A549/DDP cells, as opposed to that in A549 cells and BEAS-2B cells. (C,D) IHC results found that protein expression levels of EGR4 and OCT4, respectively, were notably higher in DDP-resistant tissues (the scale bars in IHC represent 50 μm) (left: post-DDP therapy; right: pre-DDP therapy). DDP, cisplatin; PCR, polymerase chain reaction; IHC, immunohistochemistry.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Immunohistochemistry

    Knockdown of ZNF205-AS1 led to elevated miRNA-138-5p levels in A549/DDP cells. (A) The down-regulation of ZNF205-AS1 was realized by transfecting three designed small interfering RNAs. ZNF205-AS1 level was significantly higher in blank control group and NC group than the three paralleled knockdown group (***, P<0.001). (B) The levels of miR-138-5p in A549/DDP cells with ZNF205-AS1 loss-of-function were assessed in qPCR. ZNF205-AS1 loss-of-function led to significantly up-regulation of miR-138-5p as opposed to NC group (**, P<0.01) (GAPDH and U6 were used as endogenous control for detection of ZNF205-AS1 and miRNA-138-5p, respectively). (C) The binding site of lncRNA ZNF205-AS1 with miRNA-138-5p was predicted in bioinformatics analysis. (D) The significantly inhibited relative luciferase activity of cells co-transfected with wild-type ZNF205-AS1 and miRNA-138-5p mimic, as opposed to that transfected with wild-type ZNF205-AS1 and miR-NC was detected (**, P<0.01). NC, negative control; WT, wide type; Mut, mutant; DDP, cisplatin; qPCR, quantitative polymerase chain reaction.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: Knockdown of ZNF205-AS1 led to elevated miRNA-138-5p levels in A549/DDP cells. (A) The down-regulation of ZNF205-AS1 was realized by transfecting three designed small interfering RNAs. ZNF205-AS1 level was significantly higher in blank control group and NC group than the three paralleled knockdown group (***, P<0.001). (B) The levels of miR-138-5p in A549/DDP cells with ZNF205-AS1 loss-of-function were assessed in qPCR. ZNF205-AS1 loss-of-function led to significantly up-regulation of miR-138-5p as opposed to NC group (**, P<0.01) (GAPDH and U6 were used as endogenous control for detection of ZNF205-AS1 and miRNA-138-5p, respectively). (C) The binding site of lncRNA ZNF205-AS1 with miRNA-138-5p was predicted in bioinformatics analysis. (D) The significantly inhibited relative luciferase activity of cells co-transfected with wild-type ZNF205-AS1 and miRNA-138-5p mimic, as opposed to that transfected with wild-type ZNF205-AS1 and miR-NC was detected (**, P<0.01). NC, negative control; WT, wide type; Mut, mutant; DDP, cisplatin; qPCR, quantitative polymerase chain reaction.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Negative Control, Mutagenesis, Real-time Polymerase Chain Reaction

    miRNA-138-5p gain-of-function enhanced the sensitivity of A549/DDP cells to DDP. (A) Overexpression of miRNA-138-5p was verified in qPCR assay to find that miRNA-138-5p level was significantly higher in miRNA-138-5p mimic group than NC group (*, P<0.05) (U6 as endogenous control). (B) Up-regulation of miR-138-5p promoted the invasion of A549/DDP cells. The cells were stained with 0.1% Giemsa stain (the scale bars represent 25 μm). (C) The results of CCK8 assay showed that the viability of cells exposed to gradient concentrations of DDP was significantly inhibited in miRNA-138-5p mimic group, compared with that of NC group (*, P<0.05). DDP, cisplatin; NC, negative control; IC 50 , half-maximal inhibitory concentration; qPCR, quantitative polymerase chain reaction; CCK8, Cell Counting Kit-8.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: miRNA-138-5p gain-of-function enhanced the sensitivity of A549/DDP cells to DDP. (A) Overexpression of miRNA-138-5p was verified in qPCR assay to find that miRNA-138-5p level was significantly higher in miRNA-138-5p mimic group than NC group (*, P<0.05) (U6 as endogenous control). (B) Up-regulation of miR-138-5p promoted the invasion of A549/DDP cells. The cells were stained with 0.1% Giemsa stain (the scale bars represent 25 μm). (C) The results of CCK8 assay showed that the viability of cells exposed to gradient concentrations of DDP was significantly inhibited in miRNA-138-5p mimic group, compared with that of NC group (*, P<0.05). DDP, cisplatin; NC, negative control; IC 50 , half-maximal inhibitory concentration; qPCR, quantitative polymerase chain reaction; CCK8, Cell Counting Kit-8.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: Over Expression, Staining, Giemsa Stain, CCK-8 Assay, Negative Control, Concentration Assay, Real-time Polymerase Chain Reaction, Cell Counting

    miR-138-5p up-regulation facilitated DDP-induced apoptosis of A549/DDP cells. (A) The cell apoptosis rate of DDP-treated A549/DDP cells with miR-138-5p overexpression was significantly increased than those without miR-138-5p overexpression (*, P<0.05). (B) The protein levels of Cleaved-caspase 3, Cleaved-PARP, Bax, Bcl2, and OCT4 were measured in western blotting (*, P<0.05). (C) RNA pulldown assay was performed to verify the potential binding of miR-138-5p to OCT4, and EGR4, respectively (lane 1: miR-NC input; lane 2: miR-138-input; lane 3: miR-NC pulldown; lane 4: miR-138-5p pulldown). NC, negative control; OCT4, octamer-binding protein 4; EGR4, early growth response 4; DDP, cisplatin.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: miR-138-5p up-regulation facilitated DDP-induced apoptosis of A549/DDP cells. (A) The cell apoptosis rate of DDP-treated A549/DDP cells with miR-138-5p overexpression was significantly increased than those without miR-138-5p overexpression (*, P<0.05). (B) The protein levels of Cleaved-caspase 3, Cleaved-PARP, Bax, Bcl2, and OCT4 were measured in western blotting (*, P<0.05). (C) RNA pulldown assay was performed to verify the potential binding of miR-138-5p to OCT4, and EGR4, respectively (lane 1: miR-NC input; lane 2: miR-138-input; lane 3: miR-NC pulldown; lane 4: miR-138-5p pulldown). NC, negative control; OCT4, octamer-binding protein 4; EGR4, early growth response 4; DDP, cisplatin.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: Over Expression, Western Blot, Binding Assay, Negative Control

    miRNA-138-5p loss-of-function diminished the pro-apoptosis activity induced by ZNF205-AS1-silencing in A549/DDP cells. (A) CCK8 assay confirmed that the cell viability of si-ZNF205-AS1 group was significantly lower than that with miR-138-5p knockdown and NC group (*, P<0.05). (B) Significantly higher protein levels of OCT4 and Bcl-2, as well as significantly lower protein levels of Cl-caspase-3, Cl-PARP and Bax were detected in si-ZNF205-AS1 with loss-of-function of miR-138-5p than that without (*, P<0.05). (C) The apoptosis rate of si-ZNF205-AS1 group was significantly higher than that of si-ZNF205-AS1 + miRNA-138-5p group and NC group (*, P<0.05). (D) The number of invasive cells was significantly higher in si-ZNF205-AS1 + miRNA-138-5p group and in NC group, in comparison with si-ZNF205-AS1 group (*, P<0.05). The cells were stained with 0.1% Giemsa stain (magnification 200×). (E) The migration ability of si-ZNF205-AS1 group being significantly lower than si-ZNF205-AS1 + miRNA-138-5p group (**, P<0.01) and NC group was validated in wound healing assay (the scratched areas from each group at 0 and 24 h post-wound were photographed and quantified on a microscope) (***, P<0.001). NC, negative control; DDP, cisplatin; CCK8, Cell Counting Kit-8.

    Journal: Journal of Thoracic Disease

    Article Title: Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway

    doi: 10.21037/jtd-23-1171

    Figure Lengend Snippet: miRNA-138-5p loss-of-function diminished the pro-apoptosis activity induced by ZNF205-AS1-silencing in A549/DDP cells. (A) CCK8 assay confirmed that the cell viability of si-ZNF205-AS1 group was significantly lower than that with miR-138-5p knockdown and NC group (*, P<0.05). (B) Significantly higher protein levels of OCT4 and Bcl-2, as well as significantly lower protein levels of Cl-caspase-3, Cl-PARP and Bax were detected in si-ZNF205-AS1 with loss-of-function of miR-138-5p than that without (*, P<0.05). (C) The apoptosis rate of si-ZNF205-AS1 group was significantly higher than that of si-ZNF205-AS1 + miRNA-138-5p group and NC group (*, P<0.05). (D) The number of invasive cells was significantly higher in si-ZNF205-AS1 + miRNA-138-5p group and in NC group, in comparison with si-ZNF205-AS1 group (*, P<0.05). The cells were stained with 0.1% Giemsa stain (magnification 200×). (E) The migration ability of si-ZNF205-AS1 group being significantly lower than si-ZNF205-AS1 + miRNA-138-5p group (**, P<0.01) and NC group was validated in wound healing assay (the scratched areas from each group at 0 and 24 h post-wound were photographed and quantified on a microscope) (***, P<0.001). NC, negative control; DDP, cisplatin; CCK8, Cell Counting Kit-8.

    Article Snippet: Human lung adenocarcinoma A549 cell line and human lung epithelial BEAS-2B cell line were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), and were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA), which was supplemented with 10% fetal bovine serum (FBS), and were maintained at the temperature of 37 ℃ in the 5% humidified CO 2 .

    Techniques: Activity Assay, CCK-8 Assay, Comparison, Staining, Giemsa Stain, Migration, Wound Healing Assay, Microscopy, Negative Control, Cell Counting

    (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial CaLu-3 cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

    Journal: Cell

    Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

    doi: 10.1016/j.cell.2023.12.026

    Figure Lengend Snippet: (A) Diagrams of the SARS-CoV-2 Omicron subvariants BA.2, BA.2.86, XBB.1.5, and FLip spikes. The location of specific mutations for BA.2.86 or XBB.1.5 relative to BA.2 in the N-terminal domain (NTD) or receptor binding domain (RBD) of the S1 subunit, or in the domain between fusion peptide (FP) and trans -membrane domain (TM) of the S2 subunit, or near the S1/S2 cleavage site is shown. The key mutations of FLip relative to XBB.1.5 are highlighted in red. (B and C) Infectivity of pseudotyped lentiviruses bearing each of the indicated Omicron subvariants spike was determined in (B) HEK293T cells stably expressing human ACE2 (293T-ACE2) or (C) human lung cell-derived epithelial CaLu-3 cells. Transfection efficiency and spike protein expression were comparable among all groups, which is shown in . Bars in (B–C) represent means ± standard error from triplicates. Significance relative to D614G was analyzed by a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 6). p values are displayed as ns p > 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

    Article Snippet: The cell lines in this study included human embryonic kidney 293T cells (ATCC CRL-11268, RRID: CVCL_1926), HEK293T cells expressing human ACE2 (BEI NR-52511, RRID: CVCL_A7UK), and human lung adenocarcinoma cell line CaLu-3 (RRID: CVCL_0609).

    Techniques: Binding Assay, Membrane, Infection, Stable Transfection, Expressing, Derivative Assay, Transfection

    (A–D) HEK293T cells were cotransfected with the indicated spikes of interest and GFP plasmids and were cocultured with 293T-ACE2 (A-B) or human lung epithelial CaLu-3 cells (C-D) for 24 h. Cell-cell fusion was imaged and GFP areas of fused cells were quantified (see ). D614G and no S were included as positive and negative control, respectively. Comparisons in extents of cell-cell fusion for each Omicron subvariant were made against D614G. Scale bars represent 150 μM. Bars in (B and D) represent means ± standard error. Dots represent three images from two biological replicates. Statistical significance relative to D614G was determined using a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 3). p values are displayed as ns p > 0.05, *p < 0.05, ***p < 0.001, and ****p < 0.0001.

    Journal: Cell

    Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

    doi: 10.1016/j.cell.2023.12.026

    Figure Lengend Snippet: (A–D) HEK293T cells were cotransfected with the indicated spikes of interest and GFP plasmids and were cocultured with 293T-ACE2 (A-B) or human lung epithelial CaLu-3 cells (C-D) for 24 h. Cell-cell fusion was imaged and GFP areas of fused cells were quantified (see ). D614G and no S were included as positive and negative control, respectively. Comparisons in extents of cell-cell fusion for each Omicron subvariant were made against D614G. Scale bars represent 150 μM. Bars in (B and D) represent means ± standard error. Dots represent three images from two biological replicates. Statistical significance relative to D614G was determined using a one-way repeated measures ANOVA with Bonferroni’s multiple testing correction (n = 3). p values are displayed as ns p > 0.05, *p < 0.05, ***p < 0.001, and ****p < 0.0001.

    Article Snippet: The cell lines in this study included human embryonic kidney 293T cells (ATCC CRL-11268, RRID: CVCL_1926), HEK293T cells expressing human ACE2 (BEI NR-52511, RRID: CVCL_A7UK), and human lung adenocarcinoma cell line CaLu-3 (RRID: CVCL_0609).

    Techniques: Negative Control

    KEY RESOURCES TABLE

    Journal: Cell

    Article Title: Immune evasion, infectivity, and fusogenicity of SARS-CoV-2 BA.2.86 and FLip variants

    doi: 10.1016/j.cell.2023.12.026

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: The cell lines in this study included human embryonic kidney 293T cells (ATCC CRL-11268, RRID: CVCL_1926), HEK293T cells expressing human ACE2 (BEI NR-52511, RRID: CVCL_A7UK), and human lung adenocarcinoma cell line CaLu-3 (RRID: CVCL_0609).

    Techniques: Infection, Recombinant, Transfection, Modification, Protease Inhibitor, Western Blot, Software, Imaging