hace2 egfp gene  (Addgene inc)

Journal: Scientific reports

Article Title: A new DNA aptamer which binds to SARS-CoV-2 spike protein and reduces pro-inflammatory response.

doi: 10.1038/s41598-024-58315-0

Figure Lengend Snippet: Figure 5. Inhibitory effect of DNA aptamers on the pro-inflammatory response induced by SARS-CoV-2 spike protein in HEK293 cells. (a) HEK293 cells were transfected with pcDNA3.1-E.V or pcDNA3.1-ACE2 over- expression levels were detected by RT-PCR and WB. GAPDH was used positive control. (b–d) To confirm the inhibition of the pro-inflammatory response of the candidate aptamer, 0.4 µg/mL of SARS-CoV-2 spike protein was bound with 1 µg/mL of DNA aptamers at 4 °C on a rotator for 30 min. The binders were then treated for 24 h in ACE2-overexpressing HEK293 cells. After incubation, mRNA expression of IL-1β (b), IL-6 (c), and TNF-α (d) were confirmed using qPCR, and cytokine levels [IL-1β (e), IL-6 (f), and TNF-α (g)] of culture medium was measured using ELISA kit. Negative control is random sequence ssDNA aptamer. The bar graph shows mRNA expression normalized to GAPDH (b,c,d) and cytokine concentration (e,f,g). (*, **, ***, and ns indicate P < 0.1, P < 0.01, P < 0.001, and not significant, respectively).

Article Snippet: The human ACE2 plasmid was obtained from Addgene (#154962)28.

Techniques: Transfection, Over Expression, Reverse Transcription Polymerase Chain Reaction, Positive Control, Inhibition, Incubation, Expressing, Enzyme-linked Immunosorbent Assay, Negative Control, Sequencing, Concentration Assay