SMART-seq of MG lineages in MNU-injured retinas

We further investigated injury-induced gene expression changes selectively in MG cell lineages. The tdTom-labeled MG and their descendant cells were isolated via FACS from uninjured or MNU-injured Glast-CreERT;R26tdTom/+ mouse retinas, which were infected with either AAV2-Ctrl Ab or AAV2-alpha Prox1, and mRNA expression patterns of individual cells were analyzed using SMART (Switching Mechanism At the 5' end of RNA Template) sequencing methods. We could identify a lineage flow from resting MG to RPC-like cells via activated MG, along with pseudo-time progression. The samples include Müller glia (MG) and their descendant cells isolated via FACS from the retinas of Glast-CreERT;R26tdTom/+ mice. Experimental conditions include uninjured and methyl-nitrosourea (MNU)-injured retinas, with additional treatment using AAV2-Ctrl Ab or AAV2-alpha Prox1 constructs to modulate Prox1 intake for MGs.