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. Author manuscript; available in PMC: 2022 Aug 10.
Published in final edited form as: Nat Chem. 2022 Jun 20;14(8):905–913. doi: 10.1038/s41557-022-00972-7

Figure 5. Light-induced translation in cells.

Figure 5

a) Relative Luciferase activity from HeLa cells transfected with differently capped GLuc-mRNAs. The mRNA was capped with the indicated cap analogue. Data of n=3 independent experiments are shown as mean values +/- SEM. Statistical significance was determined by two-tailed students t-test. Significance levels were defined as p<0.05:*, p<0.01:**, p<0.001:***. P value for 1(+ irr. cells, - irr. mRNA) versus 1(- irr. cells, - irr. mRNA) is 1.26 × 10−3. P value for 2(+ irr. cells, - irr. mRNA) versus 2(- irr. cells, - irr. mRNA) is 3.53 × 10−4. P value for 2(- irr. cells, + irr. mRNA) versus 2(- irr. cells, - irr. mRNA) is 1.52 × 10−4. b) Confocal laser scanning microscopy images of HeLa cells co-transfected with differently capped eGFP-mRNAs and cap 0-mScarlet-I mRNA. mRNAs contain m5C and m1Ψ. AG: ApppG-capped mRNA represents cap-independent translation. (0): m7GpppG capped eGFP-mRNA. 2: NPM cap-eGFP mRNA, either non-irradiated, irradiated in cells or irradiated before transfection (irradiated mRNA). The top row shows the eGFP fluorescence, the bottom row the mScarlet-I fluorescence. Scale bars are 50 µm. For all images background substraction was performed with ImageJ (30 pixels). Shown is one representative set of n=3 independent experiments. c) Flow cytometry of HeLa cells transfected with differently capped mRNAs (with cap analogues 0, 1, 2 or ApppG (AG)). Untransfected cells are set as gate for eGFP negative cells. Irradiation is indicated by LED icon. Shown is one representative measurement of n=3 independent experiments.