Fig 2: circROBO1 promotes prostate cancer enzalutamide resistance and glycolysis through circROBO1-miR-556-5p-PGK1 axis. (A-B) The cell proliferation rate of the C4-2B and 22RV1 prostate cancer cell lines was decreased after the inhibition of circROBO1, which was reversed after the transfection of miR-556-5p mimics. (C-D) The enzalutamide resistance was also reversed by the transfection of miR-556-5p mimics when circROBO1 was inhibited in C4-2B and 22RV1 prostate cancer cells. (E-F) The glycolysis activity of C4-2B and 22RV1 prostate cancer cells was decreased after silencing of circROBO1, which was also reversed after supplement of miR-556-5p mimics. (G) Western blot assay revealed that PGK1 expression was decreased after the suppression of circROBO1 in C4-2B and 22RV1 prostate cancer cell lines. (H) Western blot assay revealed that the decrease of PGK1 was rescued via the supplement of miR-335-5p mimics in C4-2B prostate cancer cells, which further determined the circROBO1-miR-556-5p-PGK1 axis in prostate cancer.

Fig 3: The glycolic enzyme PGK1 protein decreased in HEK293 cells expressing 3X‐TDP‐43. (A) Western blot probed for PGK1 and β‐Actin from lysed HEK293T cells transfected with CMV‐tTA and 3X‐TDP‐43 for 24 h. (B) Quantification of PGK1/β‐Actin. Unpaired T‐test with error bars representing standard deviation. ****P < 0.0001 (n = 6 biological replicates, experiment performed 3 separate times with similar results).

Fig 4: Overexpression of 3X‐TDP‐43 displays time‐dependent reduction in glycolysis enzyme protein levels. (A) Western blot probed for ADOA, PGK1, PDHK1, ENO1 normalized to β‐Actin from lysed HEK293T cells transfected with CMV‐tTA and 3X‐TDP‐43 for 24, 48, and 72 h. (B–E) Quantification of Aldolase A/β‐Actin, PGK1/β‐Actin, PDHK1/β‐Actin, and Enolase 1/β‐Actin. One‐way Anova with error bars representing standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001. Non‐significant‐ ns, P > 0.05 (n = 3 biological replicates, experiment performed 6 separate times with similar results).

Fig 5: Tau-driven spatial transcriptomic signatures precede overt tau accumulation.a NFT signature scores of genes from a snRNA dataset of AT8+ cells from patients with AD in the regions analyzed in our study. Two-tailed and unpaired t-test. b A heat map depicting the expression levels of the NFT gene set. The NFT genes are grouped into three clusters. Cluster 1: DG-specific NFT gene set; cluster 2: CA3-specific NFT gene set; cluster 3: shared NFT gene set. c The correlation between Mapt mRNA levels and NFT signature scores in WT (light green dots) and PS19 (dark green dots) ROIs. d Pgk1 gene network visualization of the gene enrichment analysis results. The color indicates the log2(fold change) of each DEG. e Pgk1 co-expressed genes based on a co-expression network analysis of RNA-seq data from AD cases and controls (Agora). Each node represents a different gene. The green shade of the circles indicates the frequency of significant co-expression; the red outline of the circles indicates that the gene is among the DEGs in our dataset. f Protein-coding genes for glycolytic enzymes in the glycolysis metabolic pathway are illustrated by BioRender.com. g A volcano plot of hit genes from CRISPR screens in the iPS cell-derived neuron. Positive hits are in red and negative in green. MAPT and PGK1 are among the top negative hits. h RNAscope staining results for Pgk1 in the CA3 region of 2-month-old WT and PS19 mice. i PGK1 is a DEG identified by large-scale human single-cell transcriptomics analysis (syn52293442 data from ROSMAP). Ast astrocytes, AG angular gyrus, braaksc.ad, non-middle versus late Braak scores (0–4 versus 5–6); braaksc.early, non-early versus middle–late Braak (0–2 versus 3–6); cogdxad, cognitive diagnosis of AD, by consensus clinical diagnosis (AD versus non-AD); EC entorhinal cortex, exc excitatory neurons, HC hippocampus, inh inhibitory neurons, mic microglia/immune cells, MT midtemporal cortex, nft NFT density in the region, nrad pathology diagnosis of AD by NIA-Reagan score, olig oligodendrocytes, PFC prefrontal cortex, plaq_d diffuse plaque density in the region, plaq_n neuritic plaque density in the region, TH anterior thalamus, vasc vascular/epithelial cells.