Fig. 7: LETR1 regulates cell proliferation and migration through transcriptional regulation of KLF4 and SEMA3C.

a Schematic representation of the experimental strategy to analyze the rescue of LETR1-ASO2 knockdown associated phenotypes with involved up- and downregulated genes after combining LETR1 knockdown and gene targets dysregulation. MOI: multiplicity of infection. b Expression levels of KLF4 after consecutive LETR1-ASO2 knockdown followed by siRNA-KD of KLF4 in neonatal LECs derived from three donors. c Quantification of the cell cycle progression analysis after 48 h consecutive knockdown of LETR1 and KLF4. Statistical analysis was performed on G0 populations. d Expression levels of SEMA3C after LETR1-ASO2 knockdown in pCDH-EV and pCDH-SEMA3C infected neonatal LECs derived from three donors. e Quantification of wound closure assay (up to 9 h) in neonatal LECs after the combination of SEMA3C overexpression and LETR1-ASO2 knockdown. Wound closure percentages were determined using TScratch¹⁰⁹. Data are displayed as mean values + SD (n = 3 in b–d; n = 10 in e). Percentages represent the knockdown efficiencies of LETR1 after the experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: not significant using RM one-way (for b, d), ordinary one-way (for c), and ordinary two-way (for e) ANOVA with Dunnett’s multiple comparisons test against scrambled control ASO—scrambled control siRNA (b), LETR1-ASO2—scrambled control siRNA (c), pCDH-EV—scrambled control ASO (d), and pCDH-EV-ASO2 (e). All displayed in vitro assays were performed in neonatal LECs derived from the same donor.