Fig. 2

Exogenous origin of Prox1 in mouse MG.

aProx1 mRNA in mouse retinal sections was visualized using RNAscope, combined with immunostaining for the MG marker Sox9. b Mean number and size of Prox1 RNAscope signals in MG nuclei and the rest INL cell nuclei are presented in the graph. Numbers of samples analyzed are shown in the graph (data from 3 independent litters). c tdTom-positive and tdTom-negative retinal cells were isolated from Glast-CreERT;R26^+/tdTom mouse retinas by FACS. Relative mRNA expression of indicated genes to Actinb1 was determined by their cycle threshold (Ct) scores obtained using RT-qPCR (data from 4 independent batches). d, e Complementary EGFP expression from the mouse Prox1 gene locus, following Cre-dependent excision of Prox1 cDNA, was assessed by co-immunostaining mouse retinal sections with chicken anti-GFP and rabbit anti-Prox1 antibodies. Cre activity in the same retinal areas was verified by expression of R26^tdTom reporter. tdTom-positive cell bodies are outlined by dotted-lines. pA, polyA transcription terminator. f EGFP intensity in tdTom-positive cells was measured by confocal microscopy. The relative intensity, normalized to the EGFP intensity of cells in Prox1^fg/+;Dkk3-Cre mouse retinas (data shown in Supplementary Fig. 3a), is plotted. Numbers of samples analyzed are shown in the graph (data collected from 3 independent litters). The mean values are indicated in the graph. g Relative Prox1 immunofluorescent intensity in MG in the corresponding retina, normalized to Prox1 intensity in BCs within the same image, is plotted. Each dot represents the median intensity collected from one retina. Numbers of samples analyzed are shown in the graph (data from 3 independent litters). P-values in the graphs were calculated using one-sided Student’s t-test (n.s., p > 0.05; ***, p < 0.005).