Journal: Cell Reports

Article Title: Drug repurposing screens reveal cell-type-specific entry pathways and FDA-approved drugs active against SARS-Cov-2

doi: 10.1016/j.celrep.2021.108959

Figure Lengend Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir. Data represent the average of three independent experiments ± SD. (C) POC for percentage of infection of the Huh7.5 drug screen performed at 0.5 μM. 33 drugs had >60% reduction in infection with >80% cell viability. (D) Distribution of 23 validated antivirals by drug target class. (E) Dose-response analysis of the candidates with a selectivity index (SI) > 3 identified in the screen. Data represent the average of three independent experiments ± SD.

Article Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir.

Techniques: High Throughput Screening Assay, Infection, Microscopy

Journal: Cell Reports

Article Title: Drug repurposing screens reveal cell-type-specific entry pathways and FDA-approved drugs active against SARS-Cov-2

doi: 10.1016/j.celrep.2021.108959

Figure Lengend Snippet: Cell-type-specific dependencies of entry inhibitors (A) Calu-3 human lung epithelial cells were infected with SARS-CoV-2 (MOI = 0.5) and processed for microscopy 48 hpi. (B) Dose-response analysis of Calu-3 cells treated with quinolines or remdesivir. Data represent the average of four independent experiments ± SD. (C) IC 50 , CC50, and SI for Vero, Huh7.5, and Calu-3 cells treated with a panel of quinolines or remdesivir. Data represent the average of four independent experiments ± SD. (D) Dose-response analysis of Calu-3 cells treated with the cathepsin inhibitor Z-FA-FMK. Data represent the average of four independent experiments ± SD. (E) Dose response analysis of Calu-3, Vero, and Huh7.5 cells treated with camostat. Data represent the average of ≥2 independent experiments ± SD. (F) IC 50 , CC50, and SI for camostat across cell types. (G) Immunoblot of Vero, Huh7.5, and Calu-3 cells probed for Ace2 and tubulin as a loading control. Representative blot is shown. (H) qRT-PCR of Ace2 or TMPRSS2 comparing Huh7.5 and Calu-3 cells. Data represent the mean ± SEM for ≥2 independent experiments.

Article Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir.

Techniques: Infection, Microscopy, Western Blot, Quantitative RT-PCR

Journal: Cell Reports

Article Title: Drug repurposing screens reveal cell-type-specific entry pathways and FDA-approved drugs active against SARS-Cov-2

doi: 10.1016/j.celrep.2021.108959

Figure Lengend Snippet: Validation of antiviral activity of nine candidates in Calu-3 cells (A) Dose-response analysis of Huh7.5 candidate antivirals in Calu-3 cells with a SI > 3. Data represent the average of ≥4 independent experiments ± SD. (B) qRT-PCR analysis of nine candidate antivirals in Calu-3 cells. Data represent the mean ± SEM for ≥2 independent experiments.

Article Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir.

Techniques: Activity Assay, Quantitative RT-PCR

Journal: Cell Reports

Article Title: Drug repurposing screens reveal cell-type-specific entry pathways and FDA-approved drugs active against SARS-Cov-2

doi: 10.1016/j.celrep.2021.108959

Figure Lengend Snippet: Cyclosporine is antiviral against SARS-CoV-2 independent of calcineurin (A) Dose-response analysis of Huh7.5 cells treated with a panel of cyclosporins and related drugs. Data represent the average of ≥2 independent experiments ± SD. (B) Dose-response analysis of Calu-3 cells treated with a panel of cyclosporins and related drugs. Data represent the average of five independent experiments ± SD. (C) Dose-response analysis of Calu3 cells treated with cyclophilin-selective drugs NIM811 and alisporivir. Data represent the average of five independent experiments ± SD. (D) Table of IC 50 s, CC50s, and SI of Huh7.5 cells and Calu-3 cells treated with the indicated drugs. (E) qRT-PCR analysis of viral replication in Huh7.5 and Calu-3 cells treated with the indicated drugs. (F) qRT-PCR analysis of viral replication in iAT2 cells treated with the indicated drugs. (G) qRT-PCR of NHBE cells treated with the indicated drug. For (E)–(G), data represent the mean ± SEM for ≥2 independent experiments.

Article Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir.

Techniques: Quantitative RT-PCR

Journal: Cell Reports

Article Title: Drug repurposing screens reveal cell-type-specific entry pathways and FDA-approved drugs active against SARS-Cov-2

doi: 10.1016/j.celrep.2021.108959

Figure Lengend Snippet:

Article Snippet: High-throughput screening in human Huh7.5 cells to identify antivirals against SARS-CoV-2 (A) Huh7.5 cells were infected with SARS-CoV-2 (MOI = 1) and 30 hpi processed for microscopy. (B) Dose-response analysis of Huh7.5 cells treated with hydroxychloroquine or remdesivir.

Techniques: Recombinant, SYBR Green Assay, Software

Journal: Molecular Therapy Oncolytics

Article Title: Intratumoral delivery of a novel oncolytic adenovirus encoding human antibody against PD-1 elicits enhanced antitumor efficacy

doi: 10.1016/j.omto.2022.04.007

Figure Lengend Snippet: AdC68-spE1A-αPD-1 induced tumor cell death in vitro (A–I) HFL-1 (A), HCT-8 (B), A549 (C), Siha (D), NCI-H508 (E), Huh7 (F), MC38 (G), CT26.WT (H), and YUMM5.2 (I) were infected with adenoviruses at indicated MOIs for 72 h. The viability of different cells was measured by CCK-8 assay. Data are shown as mean ± SEM.

Article Snippet: Human embryonic kidney (HEK) 293 cells, human normal lung fibroblast cell line (HFL-1), human colon adenocarcinoma cell line (HCT-8), human colorectal carcinoma cell line (NCI-H508), human hepatocarcinoma cell line (Huh7), and murine melanoma cell line (YUMM5.2) were purchased from ATCC (Manassas, VA, USA).

Techniques: In Vitro, Infection, CCK-8 Assay

Journal: Molecular Therapy Oncolytics

Article Title: Intratumoral delivery of a novel oncolytic adenovirus encoding human antibody against PD-1 elicits enhanced antitumor efficacy

doi: 10.1016/j.omto.2022.04.007

Figure Lengend Snippet: AdC68-spE1-αPD-1 induced tumor cell death by activating apoptotic and autophagic pathways NCI-H508 and Huh7 cells were infected with the indicated adenoviruses at 20 MOIs. The cells were collected at 24 and 48 hpi to conduct the western blot assay under denaturing condition. The proteins p62, LC3 Ι / Π , cleaved caspase-3, cleaved PARP, p53, p21, Mcl-1, CyclinD1, and CyclinE1 were analyzed. Actin was used as a loading control.

Article Snippet: Human embryonic kidney (HEK) 293 cells, human normal lung fibroblast cell line (HFL-1), human colon adenocarcinoma cell line (HCT-8), human colorectal carcinoma cell line (NCI-H508), human hepatocarcinoma cell line (Huh7), and murine melanoma cell line (YUMM5.2) were purchased from ATCC (Manassas, VA, USA).

Techniques: Infection, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: OPN promotes the Warburg effect in HCC cells. a The knockdown efficiency of OPN in HCC-LM3 cells was measured by Western blotting and ELISA. b Effects of OPN knockdown on the glucose uptake and lactate production in HCC-LM3 cells ( n = 3). c The extracellular acidification rate (ECAR) in sh-OPN and sh-Ctrl HCC-LM3 cells was measured by Seahorse analyzer ( n = 5). d Effects of OPN blockade on the glucose uptake and lactate production in HCC-LM3 cells ( n = 3). e The overexpression efficiency of OPN in NIH3T3 cells and MEFs was measured by Western blotting. f Effects of OPN overexpression on the glucose uptake and lactate production in NIH3T3 cells and MEFs ( n = 3). g Effects of OPN overexpression on ECAR in NIH3T3 cells and MEFs were measured by Seahorse analyzer ( n = 5). * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Certification of the negative regulators of HCC glycolysis. a Western blotting showed the overexpression efficiency of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, and IYD in Huh7 cells. b Real-time qPCR analysis showed the overexpression efficiency of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, and IYD in Huh7 cells ( n = 3). c-e Measurement of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, or IYD overexpression on the glucose utilization ( f , n = 3), lactate production ( g , n = 3) and ECAR ( h , n = 5) in Huh7 cells. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Western Blot, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Effects of glycolysis-related genes on HCC tumor growth. a Colony formation assay showed that OPN knockdown or blockade inhibits HCC-LM3 cell proliferation ( n = 3). b Colony formation assay for Huh3B cells treated with recombinant OPN protein ( n = 3). c Colony formation assay showed that SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, or IYD overexpression inhibits Huh7 cell proliferation ( n = 3). d The effects of glycolysis-related genes on HCC tumor growth in the presence or absence of 5 mM 2-DG ( n = 3). e In the culture medium containing 25 mM glucose or galactose, the effects of glycolysis-related genes on HCC tumor growth were analyzed by clonogenic assay. * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Colony Assay, Knockdown, Recombinant, Over Expression, Clonogenic Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: OPN promotes HCC glycolysis by modulating αvβ3-NF-κB signaling. a Blockade of integrin αvβ3 with Cilengitide inhibits glucose utilization ( n = 3), lactate production ( n = 3) and ECAR ( n = 5) in HCC-LM3 cells. b Western blotting analysis the signaling pathway influenced by OPN. c Glucose utilization and lactate production in Hep3B cells upon treatment with OPN recombinant protein and indicated pathway inhibitors ( n = 3). d Effect of OPN on the NF-κB activity in HCC cells ( n = 3). e Effect of CA-IKKβ on the glucose uptake and lactate production in OPN-silenced HCC-LM3 cells ( n = 3). * P < 0.05 and ** P < 0.01; ns: not significant

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Western Blot, Recombinant, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Inhibition of OPN-αvβ3 axis suppresses HCC tumor growth and glycolyis. a Tumor volume in sh-Ctrl and sh-OPN HCC-LM3 xenografts as indicated time point was measured ( n = 5). b Effect of Cilengitide treatment on the tumor growth of HCC-LM3 xenografts ( n = 5). c The lactate level in the tumor tissues from ( a ) and ( b ) was detected ( n = 5). d The expression of glycolytic genes in the tumor tissues from ( a ) and ( b ) was analyzed by real-time qPCR ( n = 5). e Hematoxylin and eosin staining in liver tissue samples from tumor-bearing WT and OPN-KO mice. f The expression of glycolytic genes in liver tissue samples from tumor-bearing WT and OPN-KO mice was analyzed by real-time qPCR ( n = 5). * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Inhibition, Expressing, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Expression pattern of OPN in clinical samples. a The expression of glycolytic genes in human HCC tissue samples with high OPN ( n = 10) and low OPN ( n = 20) expression was analyzed by real-time qPCR. b Representative photographs of OPN expression in HCC tumor tissues; scale bar: 50 μm. The correlation between OPN expression and the SUVmax value was analyzed. * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Expressing

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (B and C) HepG2.2.15 cells and HepG2 cells at the same cellular concentration (1 × 10 6 cells) were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively. (E and F) After the gradient transfection of pBlue-HBV 1.3-fold plasmids into HepG2 cells, the cells were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively, with the mRNA expression level of GAPDH as a reference. (H and I) Huh7 cells were analyzed in the same way as HepG2 cells. (A, D, and G) ELISA results for HBeAg and HBsAg concentrations in cell supernatants demonstrated successful infection.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Concentration Assay, Cell Culture, Expressing, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Infection

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (A and D) HBV 1.3-fold plasmids and various concentrations of PcDNA3.1–3*Flag-QPCT were cotransfected into Huh7 and HepG2 cells and cultured for 48 h, supernatants and cells were harvested. (A and D) The overexpression of Flag-QPCT in the transfected HepG2 and Huh7 cells was detected by Western blot. (B and E) The expression levels of HBeAg and HBsAg in the supernatants were detected by ELISA. (C and F) Total RNA was extracted from the cells, and the mRNA expression of HBV pgRNA in the cells was detected by RT‒PCR, with the mRNA expression level of GAPDH as a reference. (G and H) Total DNA was extracted from the cells, and the HBV-DNA copies in Huh7 and HepG2 cells were detected by RT‒PCR.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Cell Culture, Over Expression, Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay