Journal: Cell Reports Medicine

Article Title: Ancestral library identifies conserved reprogrammable liver motif on AAV capsid

doi: 10.1016/j.xcrm.2022.100803

Figure Lengend Snippet: Library production and in vitro infectivity (A) A barcoded AAV vector library was produced either with (N = 5) or without (N = 3) addition of an exogenous assembly-activating protein (AAP). Individual dots in MA plots represent distinct barcodes, colors represent the amino acid identity at position 5. To the right of MA plots, eCDF plots with SEM depicted as horizontal error bars. (B) Results of an independent sites linear elastic net regularization approach show that addition of AAP modifies the impact of certain sites on production of vector. (C) Huh7 cells were transduced with the barcoded AAV library and DNA/RNA were isolated from those transduced cells (N = 5 per condition). Individual barcodes in MA plots are colored by identity at position 3 within the library. Extant barcoded vectors were also spiked into this transduction mixture. Fold change is plotted as bar graphs (error bars determined by bootstrapping, 1,000 replicates). (D) Regularization and linear modeling approach reveal potential similarities and differences among our positions of variation with respect to transduction and gene expression in vitro .

Article Snippet: Huh7 Cell Line , ATCC , PTA-4583.

Techniques: In Vitro, Infection, Plasmid Preparation, Produced, Transduction, Isolation, Gene Expression

Journal: Cell Reports Medicine

Article Title: Ancestral library identifies conserved reprogrammable liver motif on AAV capsid

doi: 10.1016/j.xcrm.2022.100803

Figure Lengend Snippet:

Article Snippet: Huh7 Cell Line , ATCC , PTA-4583.

Techniques: Virus, Plasmid Preparation, Recombinant, Imaging, RNA Sequencing, Software, Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Optimization of hepatocyte culture systems for modeling persistent Listeria infection. Different cell seeding conditions, MOI and Listeria strains (EGDe or 10403S) were tested to obtain optimal long-term Listeria infection of HepG2 (see Supplementary Figure S1 ), Huh7 or PMH. Infected cells were examined at day 1 (d1) and at day 3 (d3) by immunofluorescence microscopy: representative examples under optimized conditions are shown. (A) Low magnification micrographs of Huh7 cells infected with EGDe strain (MOI=1-5) or PMH infected with 10403S strain (MOI=10) for the indicated time. Images are overlays of Listeria (green) and Hoechst (blue) signals (bars: 50 µm, Huh7, or 200 µm, PMH). (B) High magnification micrographs of infected Huh7 or PMH showing Listeria (green), F-actin (red) and Hoechst (blue) signals. Bars: 5 µm. Boxed regions enlarged on the right show F-actin (top) or merged signals (bottom), highlighting actin-positive bacteria at d1 and actin-negative bacteria at d3 (bars: 1 µm). (C) Micrographs of an infected Huh7 cell at d3, showing Listeria (green), LAMP1 (red) and Hoechst (blue) signals. Arrows indicate 3 examples of LisCVs. (D) Phase contrast image of an infected PMH at d3 (bars: 10 µm). Arrows indicate 2 examples of bacteria within vacuoles, shown at a higher magnification on the right (bars: 1 µm).

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Immunofluorescence, Microscopy, Bacteria

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Transcriptional responses to long-term Listeria infection in hepatocytes. HepG2, Huh7 and PMH were infected with Listeria for 72 h. (A) Cell counts (log cells per well, mean ±SD, n=3). (B) Intracellular bacteria load expressed as log CFU per well (mean ±SD, n=3). (C) Number of significant DEGs upregulated (red bars) or downregulated (blue bars) in infected hepatocytes compared to non-infected hepatocytes (adj p <0.05; |log2 FC| > 0.5). (D, E) Venn diagram showing the intersection of upregulated (D) and downregulated (E) DEGs between HepG2, Huh7 and PMH datasets. (F) The top 5 most significant KEGG pathways associated with downregulated DEGs in each of the 4 overlapping DEG datasets. (G) The top 14 most significant IPA canonical pathways associated with the Huh7–HepG2 (left) and the Huh7–PMH (right) overlapping downregulated DEGs.

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Bacteria

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Seventeen genes are upregulated by long-term Listeria infection in all hepatocyte models.

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Ubiquitin Proteomics

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Listeria infection triggers interferon responses in HepG2 and PMH, but not in Huh7 cells. (A) IFN gene expression in non-infected (NI) and infected ( Lm EGDe or 10403S) HepG2, Huh7 and PMH cells at 72 h p.i., assessed by mean FPKM for each type I, II, or III IFN gene. (a) IFNA represents all genes coding for IFN-α subtypes; (b) Ifnz is present in the mouse but not the human genome; (c) Ifnl1 is a pseudogene in the mouse genome. “n.d.”, non detectable (FPKM < 0.3, the background level). (B) RT-qPCR analysis of IFNL1 and IFNB1 transcript levels in EGDe-infected HepG2 and Huh7 cells at 8 h, 24 h, and 72 h p.i., relative to NI cells, and normalized to YWHAZ and PPIA for HepG2 and Huh7, respectively. (C) ELISA quantification of IFN-λ1 (left) and IFN-β (right) protein secreted by EGDe-infected HepG2 or Huh7 cells. Conditioned media from EGDe-infected or NI cells was sampled at 8 h, 24 h, and 72 h p.i. Dotted grey lines: limit of quantification; “n.d.”: below the limit of detection (LoD). All NI cell conditioned media sampled concurrently with infected samples were below the LoD (not shown). (D) RT-qPCR analysis of Ifnb, Ifnl2 and Ifnl3 transcript levels in 10403S-infected PMH at 24 h and 72 h p.i., expressed relative to NI cells, and normalized to Pdk3 and Pdk1 for 24 h and 72 h time points, respectively. (E) RT-qPCR analysis of MX1 transcript levels in EGDe-infected HepG2 or Huh7 cells, and Mx1 in 10403S-infected PMH, at 24 h and 72 h p.i., relative to NI cells, and normalized to YWHAZ , PPIA , and Pdk3/Pdk1 for HepG2, Huh7, and PMH respectively. Values represent mean ± SD (n=3).

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Long-term Listeria infection downregulates expression of APP genes in hepatocytes. (A) Heatmap displaying log2 FC of APP genes significantly deregulated in infected cells compared to non-infected (NI) cells. APPs are classified according to their role in either the complement or coagulation systems or other functions. Gene symbols are indicated (corresponding gene names can be found in Supplementary Tables S3, S5, S7 ). Human C1S and SERPINA1 have several murine orthologues; FC for murine C1s1 and Serpina1a are shown. (B, C) RT-qPCR analysis of representative genes in (B) EGDe-infected HepG2 or Huh7 cells, and (C) 10403S-infected PMH, relative to NI cells. A non-differentially regulated control gene ( HPRT1 for Huh7, B2M for HepG2, and Actb for PMH) is included for each set. (D) C3 protein concentration in the conditioned media of NI or infected Huh7 cells at 72 h p.i. was measured by ELISA. All values represent mean ± SD (n=3). Statistical significance, Student’s t test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Expressing, Coagulation, Quantitative RT-PCR, Control, Protein Concentration, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Long-term Listeria infection decreases the abundance of APP in the secretome of HepG2 and Huh7 hepatocytes.

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Coagulation, Binding Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Long-term Listeria infection reduces cytokine-driven expression of APP genes in human hepatocytes. (A) Huh7 cells were stimulated with IL-6 (50 ng/mL) or IL-1β (10 ng/mL) for 24 h. Gene expression was analyzed by RT-qPCR and is expressed as fold-induction relative to mock-stimulated cells; IL-6 induced the expression of C5 , C6 , C8A, FGA , FGB and HP genes, while IL-1β induced C3 , F11 and HP genes. (B, C) Effect of infection on cytokine-induced APP gene expression in Huh7 (B) and HepG2 cells (C). Cells were infected with strain EGDe (MOI~1-5) for 48 h or remained non infected (NI). IL-6 (50 ng/mL) or IL-1β (10 ng/mL) (Huh7) or IL-6 (25 ng/mL) (HepG2) was added to infected and NI cells and transcript levels relative to stimulated NI cells were quantified by RT-qPCR 24 h later (at 72 h p.i., i.e. 24 h post-stimulation). A control gene whose expression was affected neither by infection nor cytokine stimulation is included. (B) In Huh7 cells, the control gene was YWHAZ and gene expression was normalized to PPIA ; (C) In HepG2 cells, the control gene was HMBS and gene expression was normalized to YWHAZ . All values represent mean ± SD (n=3). Statistical significance, Student’s t test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Expressing, Gene Expression, Quantitative RT-PCR, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Immunomodulatory Transcriptional Signature Associated With Persistent Listeria Infection in Hepatocytes

doi: 10.3389/fcimb.2021.761945

Figure Lengend Snippet: Long-term Listeria infection downregulates almost all genes of the cholesterol biosynthesis pathway in both human HepG2 and Huh7 cells. (A) RT-qPCR analysis of transcript levels of representative genes involved in cholesterol metabolism and homeostasis in EGDe-infected HepG2 and Huh7 cells, relative to non-infected (NI) cells. Expression was normalized to YWHAZ or PPIA for HepG2 and Huh7 cells, respectively, and a non-differentially expressed control gene ( HPRT1 for HepG2, B2M for Huh7) is included. Values represent mean ± SD (n=3). Statistical significance, Student’s t test (* p < 0.05, ** p < 0.01). (B) Diagram illustrating the de novo cholesterol biosynthesis pathway. Enzyme names are in black and their corresponding gene symbols are blue when downregulated in infected cells, otherwise they are gray. Rate limiting enzymes are preceded by a star. Intermediary molecules are shown in red. Lanosterol can be transformed into cholesterol via two different routes: the Bloch pathway (left) and the Kandutsch-Russell pathway (right). (C) Heat map showing the log2 FC obtained by RNA-seq in HepG2 and Huh7 infected versus NI cells, for each of the cholesterol synthesis enzyme coding genes listed in (B) .

Article Snippet: The human hepatocellular carcinoma cell lines HepG2 (ATCC HB-8065) and Huh7 (CLS 300156) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 2 mM L-glutamine (Sigma) and 10% fetal bovine serum (FBS, Sigma) at 37°C in a humidified 5% CO 2 atmosphere and placed at 10% CO 2 during infection assays.

Techniques: Infection, Quantitative RT-PCR, Expressing, Control, Transformation Assay, RNA Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: FGL1 expression level and viability in HepG2, Huh7, Hep3B, SNU387, SNU449, and SNU475 hepatocellular carcinoma (HCC) cell lines. ( A ) Expression of FGL1 in the six HCC cell lines, as determined through immunoblotting. Data are presented as the mean ± standard deviation (SD) (n = 3). *** p < 0.001, significantly different from HCC cells with high FGL1-expressing. ( B ) Cell viability in the six HCC cell lines after sorafenib treatment. Data are presented as the mean ± standard deviation (n = 6).

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Expressing, Western Blot, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: IC 50 of sorafenib against HCC cell lines.

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Effect of sorafenib on the cell colony forming capacity and cell death of HepG2, Huh7, Hep3B, SNU387, SNU449, and SNU475 hepatocellular carcinoma (HCC) cell lines. ( A ) Number of colonies after seeding 200 cells of each HCC cell line in the plate. ( B ) Effect of sorafenib on the colony forming potential of HCC cells. The six HCC cell lines were incubated for 7 days in the presence of 0, 2, or 5 µM sorafenib. Data are presented as the mean ± standard deviation (SD) (n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001, significantly different from 0 µM sorafenib-treated cells. ( C ): Effect of sorafenib on cell death measured by flow cytometry using Annexin V-FITC and propidium iodide (PI) staining. The graph presents mean ± SD values of three independent experiments.

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Incubation, Standard Deviation, Flow Cytometry, Staining

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Effect of sorafenib on cell death and cell proliferation factors in high FGL1- and low FGL1-expressing hepatocellular carcinoma (HCC) cell lines. ( A ): Protein expression was analyzed by western blotting 48 h after treatment with 10 μM sorafenib. Expression levels of PCNA, p-ERK/ERK, LC3-II, and cleaved PARP1 quantified in Huh7 ( B ), Hep3B ( C ), SNU387 ( D ), and SNU475 ( E ) cell lines. Data are presented as the mean ± standard deviation (SD) (n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001, significantly different from 0 µM sorafenib-treated cells.

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Expressing, Western Blot, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Effect of silencing FGL1 in Huh7 and Hep3B cells on their sorafenib sensitivity. ( A ) Expression of FGL1 in Huh7 cells 24 h after FGL1 siRNA transfection. ( B ) Colony formation of Huh7 cells following siFGL1 and/or sorafenib (2 µM) treatment. ( C ) Expression of FGL1 in Hep3B cells 24 h after FGL1 siRNA transfection. ( D ) Colony formation of Hep3B cells following siFGL1 and/or sorafenib (2 µM) treatment. Graphs present mean ± standard deviation values (n = 4). * p < 0.05, ** p < 0.01, and *** p < 0.001, significantly different from scrambled siRNA-treated cells. siFGL1 (−), treated with siScr; Sorafenib (−), treated with vehicle (DMSO); siScr, scrambled siRNA; siFGL1, FGL1 siRNA.

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Expressing, Transfection, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Effect of FGL1 knockdown on sorafenib-induced cell death in Huh7 and Hep3B cell lines. Flow cytometry results of Huh7 ( A , B ) and Hep3B ( C , D ) cells are shown. The cell death percentage was estimated using the sum of the upper left, upper right, and lower right quadrants in each dot plot. The graphs present percentage of cell death. All values are presented as the mean ± standard deviation of three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001, significantly different from scrambled siRNA-treated cells; # p < 0.05, significantly different from FGL1 siRNA-treated cells. siFGL1 (−), treated with siScr; Sorafenib (−), treated with vehicle (DMSO); siScr, scrambled siRNA; siFGL1, FGL1 siRNA.

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Knockdown, Flow Cytometry, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Effect of FGL1 knockdown on p-ERK, ERK, Beclin-1, and LC3-II in hepatocellular carcinoma (HCC) cells following sorafenib treatment. Representative western blot and data from Huh7 ( A ) and Hep3B ( B ) cells. Data are presented as the mean ± standard deviation (n = 3). * p < 0.05 and ** p < 0.01, compared to siScr-treated cells. siFGL1 (−), treated with siScr. Sorafenib (−), treated with vehicle (DMSO).

Article Snippet: The cell lines and their culture media were as follows: HepG2 (ATCC, Manassas, VA, USA), DMEM; Huh7 (KCLB, Korean Cell Line Bank, Seoul, Korea), RPMI1640; Hep3B (KCLB), DMEM; SNU387 (KCLB), RPMI1640; SNU475 (KCLB), RPMI1640; SNU 449 (KCLB), RPMI1640.

Techniques: Knockdown, Western Blot, Standard Deviation

Journal: eLife

Article Title: ACE2: Evidence of role as entry receptor for SARS-CoV-2 and implications in comorbidities

doi: 10.7554/eLife.61390

Figure Lengend Snippet: Available validation data for the antibodies used in the studies described in this review in accordance to the pillars defined by the Antibodypedia validation initiative ( Uhlen et al., 2016 ). Provider refers to the information found in the website of the company. IB: Immunoblot. IHC: Immunohistochemistry. IF: Immunofluorescence. Enhanced validation, Supportive validation, No data available, + Positive detection, +/- Weak detection, - Absence of detection.

Article Snippet: AF933 Immunogen: 18-74aa , IB , , + Ovary, testis and kidney , + Airway and distal lung, ALI-cultured airway epithelial cells (correlation with mRNA levels), Calu-3 and Caco-2 cells ( ; ) - A549 (expected), Huh-7 cells ( ; ) .

Techniques: Biomarker Discovery, Western Blot, Immunohistochemistry, Immunofluorescence, Immunohistochemistry-IF, Transfection, Staining

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Exploring Cytotoxic mRNAs as a Novel Class of Anti-cancer Biotherapeutics

doi: 10.1016/j.omtm.2017.12.006

Figure Lengend Snippet: Comparison of the Decrease in Cell Viability Caused by SubA, DTA, and AA cmRNA in HuH7 and KB Cells Forty-eight hours post-transfection of KB and HuH7 cells with either of the cmRNAs SubA (subtilase cytotoxin, A-chain), DTA (diphtheria toxin, A-chain), AA (abrin-a, A-chain), or one of their stop cmRNAs, cell viability was assessed by quantifying the ATP content using the CellTiter-Glo Luminescence Viability Assay. Cell viability was proportional to the measured luminescence. 100 ng cmRNA were applied. Data are presented as mean in % ± SEM of cells transfected with the respective stop cmRNA (dotted line). Statistical significance for HuH7 and KB cells versus cells transfected with the respective stop cmRNA was assessed by Kruskal-Wallis test adjusted for multiple comparisons, with *p < 0.05, **p < 0.01, ***p < 0.001, and n = 3.

Article Snippet: HuH7 cells (CSC-C9441L, Creative Bioarray, Shirley, NY) were cultured in DMEM (1×) + GlutaMax supplemented with 10% fetal bovine serum (FBS) and 1% Pen/Strep.

Techniques: Transfection, Viability Assay