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. Author manuscript; available in PMC: 2023 Oct 1.
Published in final edited form as: Prog Neurobiol. 2022 Jul 14;217:102316. doi: 10.1016/j.pneurobio.2022.102316

Figure 1.

Figure 1.

Validation of ELAVL4 knock out (KO), over expression (OE), and rescue in iPSCs and iNs.

(A) Interaction sites for the primary antibody, qPCR primers, and sgRNA for ELAVL4 used in this study projected on a schematic representation of ELAVL4. (B) Overview of the cell lines and lentiviral conditions generated/used in this study. (C) Full KO of ELAVL4 mRNA in fADcorr iNs (P < 0.0001, t-test). (D) 18.6- and 25.5-fold increase in ELAVL4 mRNA upon ELAVL4 sv2 and ELAVL4 sv1 OE expression in fADcorr iNs respectively (P < 0.001 and P < 0.0001, Dunnett’s following a significant ANOVA). (E) Full KO of ELAVL4 mRNA in fAD iNs (P<0.0001, t-test). (F) 13.5- and 12.4-fold increase in ELAVL4 mRNA upon ELAVL4 sv2 and ELAVL4 sv1 OE expression in fADcorr iNs respectively (P < 0.0001, Dunnett’s following a significant ANOVA). (G) Representative image of immunocytochemistry of ELAVL4 rescue with ELAVL4-sv1 in fADcorr ELAVL4 KO iNs, stained with the nuclear DNA stain Hoechst, the neuronal marker βIII-tubulin and ELAVL4, mCHerry signal from the overexpression plasmid, scale bars: 10 μm. (H) Example western blots are shown for fADcorr iNs and fAD iNs, the pre-synaptic marker SYN1 is not significantly affected by manipulating the expression levels of ELAVL4. (I) Full KO of ELAVL4 protein in fADcorr iNs (P < 0.0001, t-test). (J) 20.8- and 19.5-fold increase in ELAVL4 protein upon ELAVL4 sv2 and ELAVL4 sv1 OE expression in fADcorr iNs respectively (P < 0.0001, Dunnett’s following a significant ANOVA). (K) Full KO of ELAVL4 protein in fAD iNs (P<0.0001, t-test). (L) 22.4- and 22.3-fold increase in ELAVL4 mRNA upon ELAVL4 sv2 and ELAVL4 sv1 OE expression in fADcorr iNs respectively (P < 0.0001, Dunnett’s following a significant ANOVA).