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. 2023 Dec 29;13(1):77. doi: 10.3390/cells13010077

Figure 5.

Figure 5

Modulation of FoxP3 pre-mRNA splicing with SSOs targeting both exon 2 and exon 7 cis-elements allowed us to obtain Tregs expressing a single splice variant. FoxP3 splice variant mRNA levels in Treg cells 96 h after transfection with (A) control nonspecific 36-mer nucleotides #Con1 & #Con2; (B) SSOs #Ins2 & #Ins7, which could induce the expression of the FL variant only; (C) SSOs #Del2 & #Ins7, which could induce the expression of the ∆2 splice variant only; (D) SSOs #Ins2 & #Del7, which could induce the expression of the ∆7 splice variant only; and (E) SSOs #Del2 & #Del7, which could induce the expression of the ∆2∆7 splice variant only. N = 4. The results are shown as the mean ± SD. * p ≤ 0.001 by Mann–Whitney U test. FL, full-length splice variant; ∆2, splice variant with deleted exon 2; ∆7, splice variant with deleted exon 7; ∆2∆7, splice variant with deleted both exon 2 and exon 7. ND, not detected. (F) Western blotting results demonstrate the induction of the ∆2 splice variant. Clone 150D is exon 2 specific, while clone 259D recognizes an epitope after exon 2 common for all FoxP3 splice variants.