Fig. 2: Effect of multivalency and spatial pattern on the sensitivity of TDF sensor.

A Schematic showing the design of TDF with side length of 17 base pairs (TDF-17), 26 base pairs (TDF-26), TDF sensor, 47 base pairs (TDF-47), TDF-1 with single probe site for BSJ, and TDF-2 with two probe sites. B Fluorescence intensity of TDF-17, TDF-26, TDF-47, and TDF sensor upon addition of different concentrations of circSCMH1 from plasmid-transfected HEK293T cells. Data are presented as mean ± SEM, n = 3 different measurements/group. ^***P < 0.001 versus TDF sensor group using unpaired two-tailed Student’s t test. C Fluorescence intensity of TDF-1, TDF-2, and TDF sensor upon addition of different concentrations of circSCMH1 from plasmid-transfected HEK293T cells. Data are presented as mean ± SEM, n = 3 different measurements/group. ^***P < 0.001 versus TDF sensor group using unpaired two-tailed Student’s t test. D Fluorescence intensity of TDF sensor upon addition of total RNAs and RNase R-treated total RNAs from HT-22 cells. Data are presented as mean ± SEM, n = 3 different measurements/group. P = 0.4681 versus Con group using unpaired two-tailed Student’s t test. E Fluorescence intensity of circSCMH1 from HT-22 cells treated with overexpression (OE) and knockdown (KD) of linear Scmh1 mRNAs. Data are presented as mean ± SEM, n = 3 different biological replicates/group. P = 0.9642 (Scmh1 OE) and P = 0.9657 (Scmh1 KD) versus Con group using one-way ANOVA followed by Holm–Sidak post hoc multiple comparison test. F Images of circSCMH1 in HT-22 cells after Vector (negative control), Scmh1 OE, Scmh1 KD, circSCMH1 OE, circSCMH1-∆BSJ OE and circSCMH1 KD treatment. n = 5 biological replicates/group. Scale bar, 20 μm. G Quantification of circSCMH1 intensity in HT-22 cells. Data are presented as mean ± SEM. ^***P < 0.001 versus Vector group using one-way ANOVA followed by Holm–Sidak post hoc multiple comparison test. arb. units, arbitrary units; Con, control; BSJ, back-splice junction. Source data are provided as a Source Data file.