https://orcid.org/0009-0006-0607-1872
Figures & data
Figure 1. The expression of NRSN2, cell proliferation and autophagy of HPV-transfected LC cells were enhanced.
The cells stably expressing HPV16E7 (TU212/HPV and TU138/HPV) were constructed. (a) immunoblotting was applied to assess the protein levels of HPV16E7. RT-qPCR (b), immunoblotting (c) were applied to assess the mRNA and protein levels of NRSN2. The cell viability (d), proliferation (e) were tested by CCK-8 assay and EdU staining. (F) The autophagosome was observed by TEM. (G) The protein levels of LC3B and p62 were tested using immunoblotting. **p <.01, ***p <.001.z
Figure 2. Inhibition of NRSN2 expression restrained the autophagy and malignant behavior of HPV-transfected LC cells.
knocking down NRSN2 in TU212/HPV and TU138/HPV cells. RT-qPCR (a), immunoblotting (b) were applied to test the mRNA and protein levels of NRSN2. (c) The protein levels of LC3B and p62 were tested using immunoblotting. (d) The autophagosome was observed by TEM. (e) IF was applied to assess the expression of LC3B. The cell viability (f), proliferation (g), migration (h), and invasion (i) abilities were tested by CCK-8 assay, EdU staining, wound healing assay, and Transwell assay, respectively. (j) The apoptosis was assessed using flow cytometry. *p <.05, **p <.01, ***p <.001.
Figure 3. Overexpression of NRSN2 upregulates autophagy by activating AMPK/ULK1 pathway.
Overexpression of NRSN2 and inhibition of AMPK/ULK1 using SBI (an ULK1 inhibitor) and Com (an AMPK inhibitor) in TU212/HPV and TU138/HPV cells were performed. (a) The mRNA level of NRSN2 was tested using RT-qPCR. (b) The protein level of NRSN2, p-AMPK(T172), and p-ULK1(S555) were assessed using immunoblotting. (c) IF was applied to assess the expression of LC3B. ***p <.001.
Figure 4. Knockdown of NRSN2 inhibits autophagy by suppressing AMPK/ULK1 pathway, thereby restraining the malignant behavior of HPV-transfected LC cells.
Knockdown of NRSN2 and inhibition of autophagy using RAP in TU212/HPV and TU138/HPV cells were executed. (a) The protein level of NRSN2 was assessed using immunoblotting. (b) The expression of LC3B was tested using IF. The cell viability (c), migration (d), and invasion (e) abilities were tested by CCK-8 assay, wound healing assay, and Transwell assay, respectively. (f) The apoptosis was assessed using flow cytometry. *p <.05, **p <.01, ***p <.001.
