Extended Data Fig. 2: Characterization of TDP-43 and miRNA interactions.
From: TDP-43 aggregation induced by oxidative stress causes global mitochondrial imbalance in ALS
a, Western blotting analysis of TDP-43 immunoprecipitation in N2a cells by probing TDP-43, Ago2 and GAPDH. b, Reproducibility of duplicated small RNA-seq libraries generated with total input (left) or TDP-43 IPed (right) miRNAs. c, Profile of total miRNAs versus those enriched with TDP-43 IP. Heat map show the expression miRNAs levels (left) and fold-enrichment compared to Input (right). d, Mass spectrometric analysis of proteins pulled down with biotin-labeled let-7c, biotin-labeled control RNA and naked beads without RNA bait. The data are presented in a triangular plot with yellow dots highlighting let-7c enriched proteins that include TDP-43 (top), which was further validated by Western blotting (bottom). e, RT-qPCR analysis of miRNAs enriched by myc-tag IP from N2a cells expressing myc-tagged full-length TDP-43 versus RRM1 and 2-deleted mutant. Red and Gray: target and non-target miRNAs analyzed. f, Biotin-let-7c capture of individually expressed wt and RRM1 or RRM2 deleted TDP-43 in N2a cells. Data in e (lower panel) are shown as mean ± s.d. (n = 3 independent experiments). For western blot in a and d-f, a representative example of three independent experiments is shown. Data for graph in e are available as source data online. Uncropped images for panels a and d-f are available as source data online.
