Fig. 1

TRIM22 inhibits Sp1-driven transcription. a 293T cells were seeded at 2.5 × 10⁵ cells/ml in 96-well plates. 24 h post-seeding, 0.5 ng of the minimal LTR tetO Luc reporter (WT), two deletion mutants containing either two (ΔSp1-III) or one (ΔSp1-III+II) Sp1 binding sites and a mutant with a scrambled Sp1 sequence were co-transfected in 293T cells together with rtTA-V10 encoding plasmid (0.01 ng) [11] and either TRIM22-expressing or empty control pcDNA3.1+ plasmids (5 ng). Transfected cells were cultured with doxycycline (1 µg/ml). Dual-Glo Luciferase System (Promega) was used to determine the Firefly Luc activity 48 h post-transfection according to the manufacturer’s instructions. The mean of five independent experiments ± SEM is reported. p values were calculated using two-ways ANOVA. b Either tetO-CMV or tetO-CMV + Sp1 were co-transfected in 293T cells together with a TRIM22-expressing plasmid, or a pcDNA3.1(+) plasmid as a control. rtTA-V10 encoding plasmid was co-transfected and the cells were cultured with doxycycline. Luc activity was assessed 48 h post-transfection by a Luc assay. The mean of three independent experiments ± SEM is shown. The p values were calculated using the two-ways ANOVA