Supplementary Figure 7: Analysis of Lmx1a/b-mediated transcriptional effects and gain- and loss-of-function experiments.
From: Dopaminergic control of autophagic-lysosomal function implicates Lmx1b in Parkinson's disease
(a) qRT-PCR expression analysis of genes encoding for neuronal and dopaminergic markers, and for dopaminergic transcription factors in dissected ventral midbrain region of E14.5 cLmx1a/bCtrl (cCtrl) and cLmx1a/bDatCre (cDatCre) embryos. n=5 embryos per genotype. (b) Immunostaining showing Lmx1b and tyrosine hydroxylase (TH) protein expression in primary cultures from dissected ventral midbrains 6 days after lentiviral infection with lentiviruses expressing Lmx1b (LV-Lmx1b), a short-hairpin RNA with a Scrambled sequence (shScrmbl) or a sequence targeting Lmx1b (shLmx1b). Scale bar, 100μm. n= 2 independent experiments. (c) qRT-PCR expression analysis of Lmx1a and Lmx1b in primary cultures from dissected ventral midbrains 6 days after lentiviral infection with lentiviruses expressing GFP (LV-GFP) or LV-Lmx1b, and shScrmbl or shLmx1b. n= 2 biological duplicates in at least 3 independent experiments. Data is normalized to their corresponding controls (either LV-GFP or shScrmbl). (d) qRT-PCR expression analysis of Lmx1a, Lmx1b, genes encoding for autophagic-lysosomal proteins, genes involved in intracellular transport, general neuronal and dopaminergic genes in primary cultures from dissected ventral midbrains 6 days after lentiviral infection with lentiviruses expressing RFP (LV-RFP) or Lmx1a (LV-Lmx1a). n= 3 biological duplicates. Data is normalized to the corresponding control LV-RFP. All data is represented as mean±sem of the fold change normalized against Rpl19 levels; Mann Whitney test *p<0.05, *** p<0.0005.
