Fig. 5

Chemogenetic control of endogenous protein degradation in hESC-derived cells. a, b WB analysis of EBs differentiated from SAC1 (a) and RANGAP1 (b) degron hESCs. Blotted for miniIAA7 and α-SMA. WT: wild-type; pico: 0.5 μM pico_cvxIAA treatment. c Graph showing PI4P staining intensity in EBs differentiated from SAC1 degron hESCs. Student’s t test, ****p < 0.001. All statistical comparisons are shown in Additional file 4: Table S6. d Representative images from live-imaging of cell death in EB differentiated from wild-type, RANGAP1, and SAC1 degron hESCs after 83 h of pico_cvxIAA induction. Scale bar: 50 μM. WT: wild-type; pico: 0.5 μM pico_cvxIAA treatment. e–h WB analysis in neurons differentiated from degron hESCs targeting SAC1 (e), NUP93 (f), RANGAP1 (g), and PEX3 (h). WT: wild-type; pico: 0.5 μM pico_cvxIAA treatment. i, j WB analysis (i) and immunofluorescence staining (j) of peroxisomal membrane protein PMP70 in neurons differentiated from wild-type and PEX3 degron hESCs. Scale bar: 10 μM. k Graph showing the PI4P staining intensity in neurons differentiated from wild-type and SAC1 degron hESCs. a.u. arbitrary unit. N = 20 fields. l Representative images of live-cell imaging of cell death in neurons differentiated from wild-type, RANGAP1, and NUP93 degron hESCs. Scale bar: 20 μM. Representative of 1 (RANGAP1 and NUP93) and 2 (SAC1 and PEX3) cell clones