normal cell lines a549  (ATCC)

Journal: Thoracic Cancer

Article Title: Exosomal PVRL4 Promotes Lung Adenocarcinoma Progression by Enhancing the Generation of Myeloid‐Derived Suppressor Cell‐Secreted TGF ‐β1

doi: 10.1111/1759-7714.15495

Figure Lengend Snippet: PVRL4 silencing suppresses LUAD cell proliferation, invasion, and migration and induces cell apoptosis. (A‐G) The sh‐PVRL4 or sh‐NC was transfected into A549 and HCC827 cells. (A) Western blotting for PVRL4 levels in cells. (B‐D) CCK‐8 and colony formation assays for cell proliferation. (E) Transwell for cell invasion. (F) Wound healing assay for cell migration. (G) Flow cytometry analysis for cell apoptosis. * p < 0.05.

Article Snippet: LUAD cell lines A549 and HCC827 and the normal 16HBE cells were purchased from Procell (Wuhan, China) and then cultured in RPMI‐1640 medium (PM150110) plus1% penicillin/streptomycin (PB180120) and 10% FBS (164210‐50) (Procell) at 37°C with 5% CO 2 .

Techniques: Migration, Transfection, Western Blot, CCK-8 Assay, Wound Healing Assay, Flow Cytometry

Journal: Thoracic Cancer

Article Title: Exosomal PVRL4 Promotes Lung Adenocarcinoma Progression by Enhancing the Generation of Myeloid‐Derived Suppressor Cell‐Secreted TGF ‐β1

doi: 10.1111/1759-7714.15495

Figure Lengend Snippet: PVRL4 is transferred into PBMCs from LUAD cells by exosomes. (A) TEM analysis for exosome morphology. (B) Western blotting analysis for exosomal markers (CD9, CD63 and Alix). (C) The uptake of A549‐Exo and HCC827‐Exo was observed using the PKH67 staining. (D, E) Levels of PVRL4 protein were detected by western blotting in PMSCs after incubating with A549‐Exo, HCC827‐Exo or PBS (Control). (F, G) PBMCs were incubated with PBS, A549 or HCC827 cells and PBS, or A549 or HCC827 cells and GW4869, and levels of PVRL4 protein were examined by western blotting in PMSCs. * p < 0.05.

Article Snippet: LUAD cell lines A549 and HCC827 and the normal 16HBE cells were purchased from Procell (Wuhan, China) and then cultured in RPMI‐1640 medium (PM150110) plus1% penicillin/streptomycin (PB180120) and 10% FBS (164210‐50) (Procell) at 37°C with 5% CO 2 .

Techniques: Western Blot, Staining, Control, Incubation

Journal: Thoracic Cancer

Article Title: Exosomal PVRL4 Promotes Lung Adenocarcinoma Progression by Enhancing the Generation of Myeloid‐Derived Suppressor Cell‐Secreted TGF ‐β1

doi: 10.1111/1759-7714.15495

Figure Lengend Snippet: PVRL4 knockdown in LUAD cell exosomes suppresses MDSC induction and the level of MDSC‐secreted TGF‐β1. (A‐G) PBMCs were incubated with PBS, HCC827‐Exo sh–NC , A549‐Exo sh–NC , HCC827‐Exo sh–PVRL4 , or A549‐Exo sh–PVRL4 . (A) Levels of PVRL4 were detected by western blotting. (B, C) Flow cytometry for the number of CD14 + HLA‐DR − MDSCs after co‐incubation. (D‐G) ELISA analysis and western blotting for TGF‐β1 levels in PBMCs after co‐incubation. * p < 0.05.

Article Snippet: LUAD cell lines A549 and HCC827 and the normal 16HBE cells were purchased from Procell (Wuhan, China) and then cultured in RPMI‐1640 medium (PM150110) plus1% penicillin/streptomycin (PB180120) and 10% FBS (164210‐50) (Procell) at 37°C with 5% CO 2 .

Techniques: Knockdown, Incubation, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Thoracic Cancer

Article Title: Exosomal PVRL4 Promotes Lung Adenocarcinoma Progression by Enhancing the Generation of Myeloid‐Derived Suppressor Cell‐Secreted TGF ‐β1

doi: 10.1111/1759-7714.15495

Figure Lengend Snippet: Knockdown of exosomal PVRL4 suppresses LUAD cell proliferation, invasion, and migration and induces cell apoptosis by regulating MDSC‐secreted TGF‐β1. (A‐K) A549 and HCC827 cells were incubated with TGF‐β1 overexpressed MDSCs, followed by incubating with HCC827‐Exo sh–NC , A549‐Exo sh–NC , HCC827‐Exo sh–PVRL4 , or A549‐Exo sh–PVRL4 . (A) Levels of PVRL4 were detected by western blotting. (B–E) CCK‐8 and colony formation assays for cell proliferation. (F, G) Transwell for cell invasion. (H, I) Wound healing assay for cell migration. (J, K) Flow cytometry analysis for cell apoptosis. * p < 0.05.

Article Snippet: LUAD cell lines A549 and HCC827 and the normal 16HBE cells were purchased from Procell (Wuhan, China) and then cultured in RPMI‐1640 medium (PM150110) plus1% penicillin/streptomycin (PB180120) and 10% FBS (164210‐50) (Procell) at 37°C with 5% CO 2 .

Techniques: Knockdown, Migration, Incubation, Western Blot, CCK-8 Assay, Wound Healing Assay, Flow Cytometry

Journal: Antiviral research

Article Title: Influenza virus NS1- C/EBPβ gene regulatory complex inhibits RIG-I transcription

doi: 10.1016/j.antiviral.2020.104747

Figure Lengend Snippet: Heat map of differentially expressed genes based on PCR Array analysis. NHBE cells were transfected with vector, hC/EBPβ, control siRNA or hC/EBPβ siRNA for 24h. Cells were then mock infected or infected with PR8 (1.0 MOI) for 6 h. Gene expression were analysed by real-time RT-PCR using PCR array kit as per manufacturer protocol. Data shown are average from two independent experiments. The scale shows the level of gene expression where red corresponds to upregulation (log2 fold).

Article Snippet: Human lung epithelial cell line A549, and normal human bronchial epithelial (NHBE) cells (Lonza, Switzerland) were maintained as described ( Ranjan et al., 2010 ).

Techniques: Transfection, Plasmid Preparation, Control, Infection, Gene Expression, Quantitative RT-PCR