Journal: Molecular Therapy. Nucleic Acids

Article Title: Protection of animals against devastating RNA viruses using CRISPR-Cas13s

doi: 10.1016/j.omtn.2024.102235

Figure Lengend Snippet: Schematic workflow of the used approach in this study to inhibit FMDV infection using the CRISPR-Cas system (1) Transmission and circulation of FMD disease between wildlife-livestock ecosystems due to animal interactions. (2) Finding conserved regions among different FMDV serotypes by considering multiple criteria and designing efficient and specific crRNAs by the CaSilico tool. (3) Designed crRNAs of type VI-A/B/D were synthesized as DNA oligos, and then, each crRNA was cloned into separate crRNA cloning backbones. In addition to crRNA designing, several shRNAs were designed to compare virus knockdown by RNAi and CRISPR-Cas systems. (4) Virus propagation and titration of two FMDV serotypes (O and A) were performed on BHK-21 cells to be used for viral challenge. (5) The sgRNA screening was performed to validate potent crRNAs. (6) Two methods, RT-qPCR and TCID50 assay were used to measure viral RNA and FMDV infectivity, respectively. (7) CRISPR-Cas systems can potentially be leveraged to aid antiviral development for viral diseases in animals. Figure was created with BioRender.com .

Article Snippet: To generate crRNAs and shRNAs, spacers, shRNAs, and their reverse complementary DNA oligomers were synthesized as single-stranded DNA (ssDNA) by GenScript Biotech.

Techniques: Infection, CRISPR, Transmission Assay, Synthesized, Clone Assay, Cloning, Virus, Knockdown, Titration, Quantitative RT-PCR, TCID50 Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Protection of animals against devastating RNA viruses using CRISPR-Cas13s

doi: 10.1016/j.omtn.2024.102235

Figure Lengend Snippet: Effect of Cas13 and RNAi on FMDV inhibition, serotype O (A) Experimental procedure for evaluating Cas13s and RNAi antiviral activity in BHK-21 cells. BHK-21 cells were transfected with plasmids containing targeting and non-targeting shRNAs or Cas13 and crRNAs that target or do not target FMDV 24 h before infection. Then the supernatants were collected 24 hpi to evaluate the antiviral activity. Silencing efficiency of Lwa Cas13a (B), Psp Cas13b (C), Rfx Cas13d (D), and RNAi system (E) on FMDV virus according to RT-qPCR results. In (B) to (E), data points in the graph are five independent biological experiments performed in technical replicates, lines indicate means; error bars represent SD; t test was performed to compare each treatment to non-targeting crRNA or shRNA (green and red color of the boxes indicate p < 0.05 and p < 0.01, respectively).

Article Snippet: To generate crRNAs and shRNAs, spacers, shRNAs, and their reverse complementary DNA oligomers were synthesized as single-stranded DNA (ssDNA) by GenScript Biotech.

Techniques: Inhibition, Activity Assay, Transfection, Infection, Virus, Quantitative RT-PCR, shRNA

Journal: Oncology Letters

Article Title: p53 performs an essential role in mediating the oncogenic stimulus triggered by loss of expression of neurofibromatosis type 2 during in vitro tumor progression

doi: 10.3892/ol.2017.6445

Figure Lengend Snippet: Target sequences of four candidate shRNAs.

Article Snippet: In order to obtain optimized conditions, four candidate shRNAs (sh) targeting different NF2 gene coding sequences ( ) were designed using the siDESIGN Center tool ( http://dharmacon.gelifesciences.com ) and synthesized by GeneChem Co., Ltd..

Techniques:

Journal: Oncology Letters

Article Title: p53 performs an essential role in mediating the oncogenic stimulus triggered by loss of expression of neurofibromatosis type 2 during in vitro tumor progression

doi: 10.3892/ol.2017.6445

Figure Lengend Snippet: Lentivirus-mediated shRNAs targeting NF2 in the HCT116 p53 wt cell line. Immunofluorescence was performed to verify merlin knockdown by comparing NF2 shRNA-transfected cultures (Lv- NF2 -shRNAs) with non-transfected (Blank) or nonsense shRNA-transfected cultures (Lv-Con-shRNA). The percentage of GFP-positive (green) cells in total cell numbers (blue dots, DAPI staining) was evaluated under the fluorescence microscope to screen the efficiency of viral transfection. Scale bar=100 µm. shRNA, short hairpin RNA; NF2, neurofibromatosis type 2; p53 wt , wild-type for p53; Lv, lentivirus; GFP, green fluorescent protein; Con, control.

Article Snippet: In order to obtain optimized conditions, four candidate shRNAs (sh) targeting different NF2 gene coding sequences ( ) were designed using the siDESIGN Center tool ( http://dharmacon.gelifesciences.com ) and synthesized by GeneChem Co., Ltd..

Techniques: Immunofluorescence, Knockdown, shRNA, Transfection, Staining, Fluorescence, Microscopy, Control

Journal: Oncology Letters

Article Title: p53 performs an essential role in mediating the oncogenic stimulus triggered by loss of expression of neurofibromatosis type 2 during in vitro tumor progression

doi: 10.3892/ol.2017.6445

Figure Lengend Snippet: Effect of lentiviral shRNA-mediated merlin knockdown on the proliferation of HCT116 cell lines. At 72 h after transfection of lentiviral shRNAs, indicated HCT116 cell lines in experiment groups (sh1 and sh2) and control groups (Con) were harvested, divided and subjected to additional investigations. Knockdown efficiencies of NF2 shRNAs were assessed by reverse transcription-polymerase chain reaction and western blotting in (A) p53 +/+ and (B) p53 −/− with β-actin being used as a control. Cellular proliferation was determined by a Cell Counting Kit-8 assay in (C) p53 +/+ and (D) p53 −/− . Numbers of colonies were then evaluated 7 days after seeding. Experiments (reverse transcription-polymerase chain reaction, western blotting and Cell Counting kit-8 assay) were repeated 3 times with similar results. For colony formation assay, the experiment was performed 2 times with similar results. Con: Lv-Con-shRNA (Lv-shCon); sh: Lv- NF2 -shRNA. *P<0.05 and **P<0.01 compared with controls. sh, short hairpin; Con, control; NF2, neurofibromatosis type 2; p53 +/+ ; p53 −/− , p53-null; Lv, lentivrus.

Article Snippet: In order to obtain optimized conditions, four candidate shRNAs (sh) targeting different NF2 gene coding sequences ( ) were designed using the siDESIGN Center tool ( http://dharmacon.gelifesciences.com ) and synthesized by GeneChem Co., Ltd..

Techniques: shRNA, Knockdown, Transfection, Control, Reverse Transcription, Polymerase Chain Reaction, Western Blot, Cell Counting, Colony Assay

Journal: Oncology Letters

Article Title: p53 performs an essential role in mediating the oncogenic stimulus triggered by loss of expression of neurofibromatosis type 2 during in vitro tumor progression

doi: 10.3892/ol.2017.6445

Figure Lengend Snippet: Alteration in the cell cycle pattern of HCT116 cell lines following transfection of merlin-targeted lentiviral shRNAs. Two cell types, (A) HCT116 p53 wt and (B) p53 −/− cells, treated with control shRNA (Con) or merlin-targeted shRNAs (sh1 and sh2) for 72 h, were harvested and subjected to flow cytometric analysis of DNA content subsequent to propidium idodide staining. Representative micrographs and quantification of proportions of cells in specific cell-cycle phase (G0/G1, S or G2/M) were obtained from three independent experiments. Merlin knockdown rendered significantly reduced percentages of cells in the G1 phase (between 78.5±7.3 and 56.3±6.3 and 53.4±1.9% for sh1 and sh2, respectively) and increased percentages of cells in the S phase (between 12.8±3.9 and 29.4±3.2 and 34.2±5.9% for sh1 and sh2, respectively) in HCT116 p53 wt cells. Two similar results from three independent experiments were used to plot a histogram with deviation bars. **P<0.01 compared with controls. sh, short hairpin; P53 wt , wild-type for p53; NF2, neurofibromatosis; p53 −/− , p53-null; Con, control.

Article Snippet: In order to obtain optimized conditions, four candidate shRNAs (sh) targeting different NF2 gene coding sequences ( ) were designed using the siDESIGN Center tool ( http://dharmacon.gelifesciences.com ) and synthesized by GeneChem Co., Ltd..

Techniques: Transfection, Control, shRNA, Staining, Knockdown

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Experimental design. a Schematic representation of the areas selected in our study. b Experiment one was designed to determine the effect of Rufy3 on neuronal axons. c Experiment two was designed to determine the involvement of the Rufy3 in EBI under vivo SAH conditions. d Experiment three was designed to determine the involvement of neuroprotection through the Rufy3/Rap1 complex formation via accelerating neuronal axon repair and synaptic plasticity. e Experiment four was designed to evaluate the effect of Rufy3 expression on neurocognitive function

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Expressing

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Selection of LV-Rufy3-shRNA and dose selection of LV-Rufy3-shRNA, LV-Rufy3, and 8p-CPT under normal conditions. a Representative bands of Rufy3 expression using three types of LV-Rufy3-shRNAs under the normal condition. LV-Rufy3-shRNA2 downregulated Rufy3 expression. b Representative bands of Rufy3 expression using three doses of LV-Rufy3-shRNAs (12, 18, and 24 µl/kg) under normal conditions. c Representative bands of Rufy3 expression using three doses of LV-Rufy3 (10, 15, and 20 µl/kg) under normal conditions. f Representative bands of Rap1 expression using three doses of 8p-CPT (2, 3, and 4 mg/kg) under normal conditions. d, e, g Quantitative analysis of Rufy3 and Rap1 expressions in different groups. The normal group was used as the standard. Data are shown as mean ± SEM ( n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. LV-NC or Vehicle group; # P < 0.05, ## P < 0.01, vs. 12 µl/kg-LV-shRNA group or 10 µl/kg-LV-Rufy3

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Selection, shRNA, Expressing

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Rufy3 mRNA and protein expression levels and neuronal axon damage following SAH. a Quantitative analysis of Rufy3 mRNA level. The sham group was used as a control. b Representative band of Rufy3 detected by western blot. c Quantitative analysis of Rufy3 at different stages following SAH. The sham group was used as a control. d – f Double immunofluorescence of Rufy3, MBP, and N52 (green, Alexa Fluor 488) and neuronal marker (NeuN; red, Alexa Fluor 555), and Rufy3 mainly located in the neurons. Nuclei were stained with DAPI (blue). Scale bars = 100 μm. g – i Immunopositivity of Rufy3, MBP, and N52 in neurons. The Sham group was used as the standard. Data are shown as mean ± SEM ( n = 6). * P < 0.05, ** P < 0.01, *** P < 0.01, vs. Sham group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Expressing, Control, Western Blot, Immunofluorescence, Marker, Staining

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: The protein expression levels of Rufy3 and the state of neuronal axon under LV-shRNA and LV-Rufy3 treatments after vivo and vitro SAH. a Representative bands of Rufy3 detected by western blot under 8p-CPT, LV-shRNA and LV-Rufy3 treatments following vivo SAH. b Representative bands of Rufy3 detected by western blot under LV-shRNA and LV-Rufy3 treatments following vitro SAH. c , d Quantitative analysis of Rufy3 in different groups following vivo and vitro SAH. The sham and control group were used as a control. e Double immunofluorescence analysis of Rufy3 (green, Alexa Fluor 488) and β-tubulin III (NeuN; red, Alexa Fluor 555); nuclei were stained with DAPI (blue). Scale bars = 32 μm. f, g Quantitative fluorescent intensity analysis of Rufy3 and β-tubulin III expressions in different groups. The sham group was used as the standard. h Quantitative analysis of the length of neuronal axon in different groups. i Double immunofluorescence of Rufy3 (green, Alexa Fluor 488) and β-tubulin III (axon; red, Alexa Fluor 555). Nuclei were stained with DAPI (blue). Scale bars = 100 μm. Data are shown as mean ± SEM ( n = 6). ** P < 0.01, ** P < 0.001 vs. Sham group; * P < 0.001 vs. Control group; # P < 0.05, ## P < 0.01 vs. LV-NC1 groups; & P < 0.05, && P < 0.01, &&& P < 0.001 vs. LV-NC2 group; $ P < 0.05 vs. LV-Rufy3 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Expressing, shRNA, Western Blot, Control, Immunofluorescence, Staining

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Interaction between Rufy3/Rap1 and protein expression levels of Rufy3, and activated Rap1 under LV-shRNA and LV-Rufy3 treatments after SAH. a Sample lysates with Rap1 antibody (IgG was used as a negative control) and Rufy3 were measured by immunoprecipitation. b Quantitative analysis of Rufy3/Rap1 under LV-shRNA and LV-Rufy3 treatments at 24 h after SAH. The sham group was used as a control. c Representative bands of Rap1-GTP and total Rap1 were detected by western blot. d – f Quantitative analysis of Rap1-GTP, total Rap1, and the ratio of Rap1-GTP/total Rap1 in different groups following SAH. The sham group was used as a control. * P < 0.05, ** P < 0.01, ** P < 0.001 vs. Sham group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. LV-NC1 groups; && P < 0.01, &&& P < 0.001 vs. LV-NC2 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Expressing, shRNA, Negative Control, Immunoprecipitation, Control, Western Blot

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Effects of LV-shRNA and LV-Rufy3 on the Rap1/Arap3/Rho/Fascin signaling axis after experimental SAH. a Representative bands of Fascin and Facin expressions. b, c Quantitative analysis of Fascin and Facin. The sham group was used as control. d Representative bands of ARAP3 and Rho expressions. e , f Quantitative analysis of ARAP3 and Rho. The sham group was used as control. g Double immunofluorescence of Fascin (green, Alexa Fluor 488) and β-tubulin III (NeuN; red, Alexa Fluor 555); nuclei were stained with DAPI (blue). Scale bars = 40 μm. h, i Quantitative fluorescent intensity analysis of Rufy3 and β-tubulin III expressions in different groups. The sham group was used as the standard. j Quantitative analysis of the length of neuronal axons in different groups. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. Sham group; # P < 0.05, ## P < 0.01 vs. LV-NC1 groups; & P < 0.05, && P < 0.01, &&& P < 0.001 vs. LV-NC2 group; $ P < 0.05 vs. LV-Rufy3 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: shRNA, Control, Immunofluorescence, Staining

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Effects of LV-shRNA and LV-Rufy3 on the Rap1/MEK/ERK/Synapsin I signaling axis after experimental SAH. a Representative bands of MEK1 and p-MEK1 expressions. b – d Quantitative analysis of MEK1, p-MEK1 and the ratio of p-MEK1/total MEK1. The sham group was used as control. e Representative bands of ERK1, and p-ERK1 expressions. f – h Quantitative analysis of ERK1, p-ERK1 and the ratio of p-ERK1/total MEK1. The sham group was used as control. i Representative bands of synapsin I and p-synapsin I expressions. j - l Quantitative analysis of synapsin I, p-synapsin I and the ratio of p-synapsin I/total synapsin I. The sham group was used as control. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. Sham group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. LV-NC1 groups; & P < 0.05, && P < 0.01, &&& P < 0.001 vs. LV-NC2 group; $ P < 0.05 vs. LV-Rufy3 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: shRNA, Control

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Effect of LV-shRNA and LV-Rufy3 on cortical cell apoptosis and degradation, brain edema, and neurological score after SAH. a Double immunofluorescence analysis of TUNEL staining (red, Alexa Fluor 555) and neuronal marker (NeuN; green, Alexa Fluor 488) was performed to assess neuronal apoptosis at 24 h after SAH. b Fluoro-Jade C staining (green) was performed to evaluate neuronal degeneration and arrows pointed to FJC-positive cells. c Quantitative analysis of apoptotic neuron percentage. d Quantitative analysis of Fluoro-Jade C positive cells/mm 2 in brain sections in each group. e Double immunofluorescence of MBP (green, Alexa Fluor 488) and neuronal marker (NeuN; red, Alexa Fluor 555), and Rufy3 mainly located in the neurons. f Brain water content. g Neurological scoring. Scale bars = 100 μm. *** P < 0.001 vs. Sham group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. LV-NC1 groups; & P < 0.05, &&& P < 0.001 vs. LV-NC2 group; $ P < 0.05, $$$ P < 0.001 vs . LV-Rufy3 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: shRNA, Immunofluorescence, TUNEL Assay, Staining, Marker

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Effect of silenced/overexpressed Rufy3 on the recovery of sensory and locomotor function of rats after SAH. a – d Adhesive removal test. Silencing Rufy3 delayed, while overexpressing Rufy3 accelerated, the recovery of sensory function in rats after SAH. e – h Rotarod test. Silencing Rufy3 delayed, while overexpressing Rufy3 accelerated, the recovery of locomotor function in rats after SAH. Data are shown as the mean ± SEM (n = 12). *** P < 0.01, vs. Sham group; # P < 0.05, ### P < 0.001, vs. LV-NC1 group, & P < 0.05, &&& P < 0.001 vs. LV-NC2 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Adhesive

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Effect of silenced/overexpressed Rufy3 on the recovery of spatial and motor learning ability of rats after SAH. a – f Swimming tracks (red) in the Morris water maze (MWM). Silencing Rufy3 had a longer trajectory, whereas overexpressing Rufy3 had a shorter trajectory. The green circle indicates the target platform. g Swimming distance in the MWM. Silencing Rufy3 increased the swimming distance, whereas overexpressing Rufy3 decreased the swimming distance. h Escape latency in the MWM. Silencing Rufy3 increased the escape latency, whereas overexpressing Rufy3 decreased the escape latency. Data are shown as mean ± SEM (n = 10). *** P < 0.01, vs. Sham group; ### P < 0.001, vs. LV-NC1 group; &&& P < 0.001, vs. LV-NC2 group

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques:

Journal: Molecular Brain

Article Title: Roles of Rufy3 in experimental subarachnoid hemorrhage-induced early brain injury via accelerating neuronal axon repair and synaptic plasticity

doi: 10.1186/s13041-022-00919-6

Figure Lengend Snippet: Schematic diagram of the potential neuroprotective mechanisms of Rufy3 after SAH. Following SAH, the inhibition of Rufy3 expression resulted in an increase in neuronal axon and synaptic damage accompanied by hindered Rufy3/Rap1 complex formation. In addition, Rufy3 overexpression contributed to Rufy3/Rap1 complex formation and accelerated neuronal axon repair via activating the Rap1/Arap3/Rho/Fascin signaling axis. It also accelerated synaptic plasticity by activating the Rap1/MEK/ERK/Synapsin I signaling axis after experimental SAH

Article Snippet: Three lentiviral Rufy3 shRNAs (83759-1, 83580-1, and 83578-1) and a negative control virus (LVCON313, LV-NC1) were purchased from Genescript (Nanjing, China).

Techniques: Inhibition, Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: Hypoxia-induced long noncoding RNA NR2F1-AS1 maintains pancreatic cancer proliferation, migration, and invasion by activating the NR2F1/AKT/mTOR axis

doi: 10.1038/s41419-022-04669-0

Figure Lengend Snippet: A Image of xenograft tumors resected from nude mice-carrying PANC-1 cells that transduced with lentiviruses encoding sh-Control, sh-NR2F1-AS1#1, or sh-NR2F1-AS1#2 ( n = 5). B Growth curves of the subcutaneous tumor were made from weekly measurements of tumor volume. C Weights of the subcutaneous tumors in each group were measured after 5 weeks when the mice were sacrificed. D Representative images of IHC staining for Ki-67 and PCNA in the tumor sections from sh-Control, sh-NR2F1-AS1#1, or sh-NR2F1-AS1#2 xenografts; scale bar = 100 μm. E – H Representative fluorescent images of microscopic metastatic nodules after 5 weeks ( n = 5), and liver ( G ) and lung ( H ) tissues stained with hematoxylin and eosin (H&E); scale bar = 20 μm. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Short hairpin RNAs (shRNAs) containing NR2F1-AS1 mimics, NC mimics, sh-Control, sh-NR2F1-AS1#1, and NR2F1-AS1#2 lentivirus were provided by Shanghai Genechem Co., Ltd. (Shanghai, China).

Techniques: Transduction, Control, Immunohistochemistry, Staining