Fig 1: Immunohistochemistry for AKT1, GTSE1, BIRC5, AURKA, PBK, KNSTRN, and PSMB10. Samples from cancer-adjacent endometrial tissue (N = 75) and endometrial cancer (N = 107). Cancer-adjacent endometrial tissue sample of weak immunostaining score for either AKT1 (A), GTSE1 (D), BIRC5 (G), AURKA (J), PBK (M), KNSTRN (P), and PSMB10 (S). Endometrial cancer sample of weak and strong immunostaining score for either AKT1 (B,C), GTSE1 (E,F), BIRC5 (H,I), AURKA (K,L), PBK (N,O), KNSTRN (Q,R), and PSMB10 (T,U). The expression for each gene were depicted in (V) slides (200 X, *p < 0.05, **p < 0.01).
Fig 2: (A) Tumor tissue and corresponding non-tumorous adjacent tissue were collected from colorectal cancer patients in TMAs. The expression of PARPBP (green), KNSTRN (red), and KIF2C (cyan) indicates the lipid droplets in tumor-infiltrating myeloid cells. Nuclei (blue) were stained using DAPI. The absolute number of positive cells was quantified in whole fields (hpf; scale bar = 20 lm). (B) Comparing the expression of PARPBP, KNSTRN, and KIF2C in tumor tissue and non-tumorous adjacent tissue based on the single index strength score (Student’s two-tailed t-tests, ***p < 0.001). (C) The Kaplan–Meier test of the risk score for the overall survival between the high-risk and the low-risk group (log-rank test, p < 0.05). (D) The calibration curves for identifying the consistency between actual observation and nomogram-predicted OS probabilities.
Fig 3: Overall (OS) and progression-free survival (PFS) curves in endometrial cancer (N = 107) according to AKT1 (A,B), GTSE1 (C,D), BIRC5 (E,F), AURKA (G,H), PBK (I,J), KNSTRN (K,L), and PSMB10 (M,N) genes expression status.
Fig 4: Astrin loss promotes anabolic pathways.(A) Volcano plot of label-free proteomics of WT and SPAG5 ind KO HeLa cells (n = 4 independent replicates). CLUH targets are marked in blue. KEGG pathways of downregulated (B) or upregulated (C) proteins (with a cutoff of p = 0.05; q = 0.15) detected in proteomics analysis of SPAG5 ind-KO cells (A and Supplementary file 3) using the EnrichR webtool. Targeted metabolomics of WT, SPAG5, and KNSTRN ind-KO cells after 8 hr Hanks' Balanced Salt Solution (HBSS) starvation showing glycolytic intermediates and lactic acid (D), sedoheptulose-7-P (E), pyruvic acid and TCA cycle intermediates (F), nucleotide levels (G), and ornithine and polyamine levels (H). Bars show mean ± standard error of the mean (SEM) and dots represent values of individual replicates. One-way analysis of variance (ANOVA) with post hoc Tukey’s multiple comparison tests on log converted fold changes were performed with *p = 0.05; **p = 0.01; ***p = 0.001.
Fig 5: Astrin and kinastrin depletion does not mimic mitochondrial phenotypes seen upon absence of CLUH.(A) Mitochondrial network of WT, SPAG5, and KNSTRN ind-KO HeLa cells grown in basal or galactose media for 16 hr. Mitochondria were stained with an antibody against TOMM20. Scale bar, 10 µm. On the left side, confocal immunofluorescence pictures are shown, on the right side, mitochondrial network is shown after segmentation. Color code for different mitochondrial morphologies: purple: filamentous; green: rod-shaped; orange: fragmented; blue: swollen; black: unclassifiable. (B) Quantification of mitochondrial morphology of experiments as shown in A (n = 3 independent experiments; at least 66 in basal or 44 cells in galactose have been analyzed per genotype per replicate). Bars show the mean of each morphological class with standard error of the mean (SEM). (C) mtDNA levels of WT, SPAG5, and KNSTRN ind-KO HeLa cells grown in basal or galactose media for 16 hr (n = 4 independent experiments). Bars show mean ± SEM and dots represent values of individual replicates. (D) Blue native polyacrylamide gel electrophoresis (BN-PAGE) analysis of respiratory chain supercomplexes and complex I activity staining of isolated mitochondria of WT, CLUH KO and WT, SPAG5, and KNSTRN ind-KO HeLa cells grown in basal or galactose media for 16 hr. Figure 4—source data 1.Uncropped blots for Figure 4D. Figure 4—source data 2.Unedited blots for Figure 4D.
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