Fig. 2: REVERSIR-mediated regulation of an erythropoietin transgene.

a Vector genome schematics for AAV encoding mouse EPO (mEPO) transgene under control of TTR shRNA with intact (self-silencing; TTR shRNA/TTR-ts) or scrambled (non-self-silencing; TTR shRNA/NT-ts) binding site in 3’ UTR. b Mice were transduced with 2 × 10¹¹ GC of AAV8 vectors shown in (a) for two weeks and then subcutaneously injected with REVERSIR. Serum was collected for quantification of mEPO levels by ELISA at indicated days prior to and following REVERSIR administration. c Serum EPO concentrations in mice that received 0.1 mg/kg 9-mer TTR or NT REVERSIR, compared to vehicle control on D0 (n = 4 mice for vehicle groups and n = 3 mice for REVERSIR groups). Data were analyzed by two-way repeated measures ANOVA with Tukey’s post hoc test (^***p < 0.001 relative to TTR shRNA/TTR-ts + NT REVERSIR control). d EPO concentrations following treatment with increasing doses of TTR REVERSIR 2 (0.01, 0.03, 0.1, or 0.3 mg/kg). Experiments in panels (c) and (d) were conducted in parallel and compared to the same vehicle conditions. An additional control was included in (d) in which mice transduced with the non-self-silencing vector were treated with TTR REVERSIR (n = 4 mice for vehicle and 0.03 mg/kg REVERSIR groups; n = 3 mice for 0.01 and 0.1 mg/kg REVERSIR groups; n = 2 mice for 0.1 and 0.3 mg/kg REVERSIR groups). Statistical testing was performed by using two-way repeated measures ANOVA followed by Tukey’s test for multiple comparisons (^*p < 0.05 relative to TTR shRNA/TTR-ts + vehicle control). #, adjusted p = 0.049 at D3 compared to TTR shRNA/TTR-ts + vehicle condition by multiple unpaired t-test (one per row) corrected for multiple comparisons by Holm-Sidak method (alpha=0.05). All error bars represent s.e.m. AAV vector and experimental design illustrations were created with BioRender.com. Source data with detailed statistical analyses are provided as a Source Data file.