Fig. 2: Lentiviral expression of SRCP1 had limited impact on insoluble protein burden in ALS iPSC-derived motor neurons. | Gene Therapy

Fig. 2: Lentiviral expression of SRCP1 had limited impact on insoluble protein burden in ALS iPSC-derived motor neurons.

From: Viral vector gene delivery of the novel chaperone protein SRCP1 to modify insoluble protein in in vitro and in vivo models of ALS

Fig. 2

Live fluorescence imaging of iPSC-derived motor neurons showed transgene expression upon infection with lenti-GFP (A) and lenti-SRCP1-RFP (B). Both SOD1 and C9orf72 ALS iPSC-derived motor neurons exhibited increased insoluble SOD1 and TDP-43 protein compared to control motor neurons, and lentiviral expression of SRCP1 did not reduce insoluble SOD1 or TDP-43 protein burden compared to lenti-GFP infected or uninfected (UTX) conditions. Lentiviral infection in healthy (control) iPSC-derived motor neurons did not alter insoluble protein burden. The lack of HSP70 expression demonstrates the efficiency of the insoluble protein purification process (C). Lentiviral expression of SRCP1 did not alter soluble SOD1 or TDP-43 protein expression in iPSC-derived motor neurons compared to lenti-GFP infected or uninfected (UTX) conditions. HSP70 protein expression across the samples indicates consistent soluble protein extraction. Data were normalized to total protein (D). Quantification of soluble protein showed clear increases in soluble TDP-43 protein burden in SOD1 and C9orf72 iPSC-derived motor neurons compared to control motor neurons. Neither soluble SOD1 nor HSP70 protein expression levels were different between ALS iPSC-derived motor neurons and control motor neurons (E). n = 5, *p < 0.01 by ANOVA. Data are graphed as mean ± 95% CI.

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