Skip to main content
. 2022 Jun 2;31(20):3478–3493. doi: 10.1093/hmg/ddac128

Figure 3.

Figure 3

OPA1 haploinsufficiency does not impair in vitro iPSC-RGC differentiation. (A–D) Temporal qPCR analysis of key RGC transcription factors PAX6, RAX, LHX2 and ATOH7 (A) and terminal RGC genes BRN3B (B), ISL1 (C) and SNCG (D) at D-1, D7, D21 and D42 in WT control and OPA1+/− differentiations. Gene expression was first normalized to the geometric mean of GAPDH and ACTIN expression, and then normalized to D-1 undifferentiated iPSC to determine fold change. Bars represent mean expression ± SEM. n = 3–4 RNA samples per time point from 3–4 independent differentiations. **P < 0.005, ***P < 0.01, ****P < 0.001. (E–G) Immunofluorescent analysis of OPA1+/− and control iPSC-RGCs at D42 to confirm expression of terminal differentiation markers. RGCs express transcription factors BRN3A and BRN3B (E), ISL1 (F) and SNCG (G) and cytoskeletal proteins neurofilament and βIII tubulin (TUBB3) (F,G). Cell nuclei identified with DAPI. Scale bars = 40 μm.