GSK3β inhibition by CHIR 99021 affects neuronal differentiation of neuroepithelium in a dose dependent manner-IHC.

A. Representative images of immune-stained organoids for detection of NPC marker PAX6, neuronal markers DCX and TUJ1, intermediate progenitor marker TBR2, and B. neuroepithelial marker E-cadherin. C. Quantification of the cell type marker positive areas normalized by the DAPI positive area for each organoid, and relative to DMSO organoids (E-Cad: DMSO: 1 ± 0.5046, n = 3; 1μM: 0.8878 ± 0.3554, n = 3; 10μM: 92.47± 11.34, n = 2; DMSO vs 1μM, **: p-value = 0.8646; DMSO vs 10μM, **: p-value = 0.0040. PAX6: DMSO: 1 ± 0.2189, n = 5; 1μm: 1.834 ± 0.1585, n = 7; 10μM: 0.5 ± 0.09146, n = 2; DMSO vs 1μM, **: p-value = 0.0099; DMSO vs 10μM, p-value = 0.2339. TBR2: DMSO: 1 ± 0.2185, n = 6; 1μM: 2.2 ± 0.7904, n = 5; 10μM: 5.5 ± 1.471, n = 5; DMSO vs 1μM, p-value = 0.1456; DMSO vs 10μM, **: p-value = 0.0087; DCX: DMSO: 1 ± 0.3027, n = 5; 1μM: 1.078 ± 0.1477, n = 7; 10μM: 0.1283± 0.2005, n = 2; DMSO vs 1μM, p-value = 0.8046; DMSO vs 10μM, *: p-value = 0.0449; TUJ1: DMSO: 1 ± 0.1175, n = 6; 1μM: 1.17 ± 0.3217, n = 5; 10μM: 1.512 ± 0.3407, n = 5; DMSO vs 1μM, p-value = 0.6026; DMSO vs 10μM, p-value = 0.1583; unpaired t-test.). Bar graphs represent mean ± SEM.