Journal: bioRxiv
Article Title: ZNF423 depletion induces the integrated stress response and represents a potential vulnerability in NF1-associated MPNST
doi: 10.64898/2026.03.03.709360
Figure Lengend Snippet: (A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, ATF3, ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.
Article Snippet: Immunoblots were performed using primary antibodies against ZNF423 (ABN410, Sigma-Aldrich), GAPDH (sc-365062, Santa Cruz Biotechnology), SUZ12 (3737S, Cell Signaling Technology), ATF3 (18665S, Cell Signaling Technology), ATF4 (11815S, Cell Signaling Technology), PARP (9542S, Cell Signaling Technology), and γH2A.x (9718S, Cell Signaling Technology).
Techniques: RNA sequencing, Control, Transfection, Two Tailed Test, Western Blot, Expressing