Enriched single-cell LHON-RGC population recapitulates LHON pathological features. (A) Bright field images showing morphologies of wild type (normal) and LHON-specific hiPSCs, EBs, NRs, OVs, and 2-day RGCs. Scale bar: 100 µm. (B) Western blot showing protein expression level of TuJ1 in OVs and RGCs derived from normal and LHON-specific hiPSCs. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) used as a loading control. (C) Electrophysiological analysis of day-21 Normal-RGCs and LHON-RGCs. Patch clamp was used to detect the spikes of action potential of cells submerged in tetrodotoxin-free artificial cerebrospinal fluid (ACSF) and ACSF containing 0.5 μM tetrodotoxin for negative control (TTX). * p < 0.05, ** p < 0.01.