Two missense STK11 gene variations impaired LKB1/adenosine monophosphate-activated protein kinase signaling in Peutz-Jeghers syndrome

Figure 3 Expression and functional analysis of STK11. A: Plasmids were successfully transfected into HeLa cells. HeLa cell-Lipofectamine 3000; and HeLa cell transfected with enhanced green fluorescent protein (EGFP)-Empty, EGFP-N1-STK11, EGFP-N1-STK11 c.889A>G (p.Arg297Gly), EGFP-N1-STK11 c.733C>T (p.Leu245Phe) and EGFP-N1-STK11 c.910C>T (p.Arg304Trp) plasmid vectors, Magnification: 10 ×; B: Plasmid transfection efficiency was high; C: PCR gel electrophoresis confirmed that the transfected plasmid was correct; D: The endogenous expression of STK11 gene was low in HeLa cells; E: The exogenous expression of STK11 gene in HeLa cells after transfection; F: These missense variants do not affect gene transcription; G: These variations do not affect the function of gene translation into proteins; H: These missense variants disrupted the phosphorylation of liver kinase B1 and adenosine monophosphate-activated protein kinase; I: These missense variants promote cell proliferation, ^cP < 0.0001. “R” represents arginine; “G” represents glycine; “L” represents leucine; “F” represents phenylalanine; and “S” represents serine. Independent sample results were obtained after at least three experiments and then obtained statistically. NS: Not significant; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; LKB1: Serine/threonine kinase 11.