Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 1 Male Slc3a1 KO mice exhibited severer stone formation and renal injuries. (A) Relative urinary cystine concentration of Slc3a1 KO and WT mice from both sexes. (B) Typical hexagonal cystine crystals from the urine of Slc3a1 KO and WT mice. Scale bar,100 mm. (C) Bladder stones in 20-week-old Slc3a1 KO mice from both sexes. (D) Serum blood urea nitrogen (BUN) and creatinine measurement of Slc3a1 KO and WT mice from both sexes. (E) Protein levels of KIM1 in the kidneys of Slc3a1 KO and WT mice from both sexes. (F) Relative mRNA levels of fibrosis and inflammatory genes (Col3a1, Vimentin, Fibronectin, IL-1b, and TNF-a) in the kidneys of Slc3a1 KO and WT mice from both sexes. (G) Representative images (left) of hematoxylin/eosin-, Sirius red-, and TUNEL- stained kidney sections of Slc3a1 KO and WT mice from both sexes and their quantification (right). Scale bar, 100 mm. All data are

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: Concentration Assay, TUNEL Assay, Staining

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 2 The male susceptibility to cystinuria is independent of Slc7a13. (A) The published single-cell RNA sequencing data representing the expression levels of Slc3a1 and Slc7a13 in mouse kidneys. The highlighted numbers are as below: 3-segment 1 of the proximal tubule (female), 4-segment 1 of the proximal tubule (male), 5-segment 2 of the proximal tubule (female), 6-segment 2 of the proximal tubule (male), 7-segment 3 of the proximal tubule (female), 8-segment 3 of the proximal tubule (male). The annotation of other numbers shown here can be found at https://cello.shinyapps.io/kidneycellexplorer/. (B) Relative mRNA levels of Slc7a9, Slc7a13, and Slc3a1 in the WT mouse kidneys. (C) Schematic diagram of the Slc7a13 and Slc3a1 double knockout (DKO)

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: RNA Sequencing, Expressing, Double Knockout

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 3 The orchidectomy procedure failed to mitigate the sexual disparity observed in cystinuria. (A) Study design of the orchiectomy procedure with Slc3a1 KO males. (B) Bladder stones from control Slc3a1 KO males and orchiectomized Slc3a1 KO males at 20 weeks old. (C) Blood urea nitrogen (BUN) measurement of WT males, sham Slc3a1 KO males, and orchiectomized Slc3a1 KO males. (D) Relative mRNA levels of fibrosis and immune markers (Col3a1, Vimentin, Fibronectin, IL-1b, and TNFa) in the kidneys of WT males, sham Slc3a1 KO males, and orchiectomized Slc3a1 KO males. (E) Representative images (left) of hematoxylin/eosin- and Sirius red-stained kidney sections of WT males, sham Slc3a1 KO males, and orchiectomized Slc3a1 KO males, and their quantifi- cation (right). Scale bar, 100 mm. All data are represented as mean standard error mean. p values were calculated by one-way ANOVA with the post hoc Tukey test. ****p < 0.0001, **p < 0.01, *p < 0.05.

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: Control, Staining

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 4 Mitochondria might serve as a central target for the observed sex differences in Slc3a1 KO mice. (A) Schematic diagram of the bulk RNA sequencing (RNA-seq) with kidneys from Slc3a1 KO females and males. (B) Graph of the differentially expressed genes in bulk RNA-seq with 1953 up-regulated (red) and 663 down-regulated genes (blue). (C) Gene set enrichment analysis (GSEA) analysis of collagen-containing extracellular matrix, NF-kB signaling, and apoptotic signaling pathways of KO male kidneys over female kidneys from bulk RNA-seq data. NES, normalized enrichment score; Pval, nominal p-value. (D) The top 15 GSEA enrichment terms with the lowest p-values comparing KO male kidneys with female kidneys from bulk RNA-seq data.

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: RNA Sequencing, Protein-Protein interactions

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 5 The male kidney of Slc3a1 KO exhibited impairments in mitochondrial functions. (A) Representative transmission electron micrographs of mitochondria in the renal tubule of Slc3a1 KO male and female mice. Scale bar, 1 mm in the upper panel and 533 nm in the lower panel. (B) Relative mRNA levels of Cox4 and OXPHOS genes (Ndufb8, Sdhb, and Uqcrc2) in the kidneys of Slc3a1 KO male and female mice. (C) Western blots of OXPHOS genes (NDUFB8, SDHB, UQCRC2, MTCO1, and ATR5A) in the kidneys of Slc3a1 KO male and female mice (upper) and the quantification of their protein levels (lower). (D) Western blots of HSP60 in the kidneys of Slc3a1 KO male and female mice. (E) Relative mtDNA levels in the kidneys of Slc3a1 KO male and female mice using quantitative PCR by amplification of ND1 and 16S genes and normalization against the hexokinase 2 (HK2) gene. (FeH) Quantifi- cation of fluorescent JC1 signaling (red/green ratio), fluorescent ATP signaling, and fluorescent mitoSOX signaling in primary cells from the kidneys of Slc3a1 KO male and female mice. All data are represented as mean standard error of the mean. p values were calculated by t-test with the post hoc Tukey test. *p < 0.05.

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: Transmission Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 6 High SLC3A1 protein level was associated with enhanced mitochondrial functions in the kidney. (A) Protein levels of SLC3A1 in the kidneys of WT and Slc3a1 KO mice from both sexes. (B) SLC3A1 expression in the kidneys of Slc3a1 KO and WT mice from both sexes by immunohistochemical staining. Scale bar, 100 mm. (C) The relative mRNA expression of SLC3A1 in the healthy human samples. (D) The content of ATP and GSH in the primary tubule cells derived from kidneys of SLC3A1high males and SLC3A1low

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: Expressing, Immunohistochemical staining, Staining, Derivative Assay

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 7 Elevated SLC3A1 protein level was associated with enhanced mitochondrial functions in proximal tubule cells. (A) Schematic diagram of the single-cell RNA sequencing (scRNA-seq) with kidneys from SLCA31high males and SLCA31low females. (B) The UMAP of 24 distinct cell types identified by unsupervised clustering. Podo, podocyte; Pe, parietal epithelium; PT-S1, S1 segment of proximal tubule; PT-S1/S2, S1 and S2 segments of proximal tubule; PT-S2/S3, S2 and S3 segments of proximal tubule; PT-S3, S3 segment of proximal tubule; LOH-j, LOH of juxtamedullary nephron; LOH-c, LOH of cortical nephron; LOH-Thin-U, thin limb of upper LOH; LOH-Thin-L, thin limb of lower LOH; LOH-Thick, thick limb of LOH; Md, macula densa; DCT, distal convoluted

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: RNA Sequencing

Journal: Genes & Diseases

Article Title: Mitochondrial SLC3A1 regulates sexual dimorphism in cystinuria

doi: 10.1016/j.gendis.2024.101472

Figure Lengend Snippet: Figure 8 SLC3A1 mediated mitochondrial functions by NAD þ uptake. (A) Schematic diagram of SLC3A1 overexpression experi- ments. (B) Transcript expression of GSH metabolism related genes Gclc and Gclm in the transfected or non-transfected cells. (C) Transcript expression of fatty acid metabolism related genes Cpt1a, Acox1, Cpt2, and Acox2 in the transfected or non-transfected cells. (D) Representative images of double staining of SLC3A1 and the mitochondrial marker ATP5A1 in HK2 cells. Scale bar, 10 mm. (E) NADþ content of control and SLC3A1 overexpressed cells. (F) NADþ content of mitochondria isolated from HEK 293T control cells and cells stably overexpressing (OE) SLC3A1 before and after a 20-min incubation with 1 mM NADþ. (G) Scheme of the experimental

Article Snippet: Immunostaining was performed using the primary antibodies against SLC3A1 (Proteintech #19912-1- AP) and ATP5A1 (Proteintech #66037-1-Ig).

Techniques: Over Expression, Expressing, Transfection, Double Staining, Marker, Control, Isolation, Stable Transfection, Incubation

Journal: Journal of Functional Foods

Article Title: Eicosapentaenoic acid and docosahexaenoic acid suppress colonic tumorigenesis in obese mice

doi: 10.1016/j.jff.2024.106164

Figure Lengend Snippet: Fig. 7. Schematic diagram of the mechanism of EPA and DHA inhibiting colon tumor in obese mice. This mechanism involves solidifying the intestinal mucosal barrier by improving the intestinal flora dysbiosis, suppressing inflammatory signaling pathways such as TNF-α/NF-κB, IL-6/STAT3, and NLRP3/IL-1β pathways, and blocking the GSK3β/β-catenin/C-myc carcinogenic pathway.

Article Snippet: Primary antibodies against NLRP3 (15101S, 1:1000), Stat3 (12640S, 1:1000), p-Stat3Tyr705 (9131S, 1:1000), ERK (9102S, 1:1000), p-ERK (4370S, 1:2000), Akt (9272S, 1:1000), p-AktSer473 (9271S, 1:1000), p-GSK3βSer9 (9336S, 1:1000), β-catenin (9562S, 1:1000), C-myc (9402S, 1:1000), Cyclin-D1 (2922S, 1:1000), PCNA (13110S, 1:1000), NF-κB (8242S, 1:1000), and horseradish peroxidase (HRP)-conjugated secondary antibodies (7074S, 1:2000) were obtained from Cell Signaling Technology, Inc. (Danvers, USA).

Techniques: Protein-Protein interactions, Blocking Assay