Fig 1: DCHS1‐Mut promotes HK‐2 cell apoptosis and autophagy. (a) Flow cytometric analysis of apoptotic cells was quantified by Annexin V‐FITC/PI double‐staining. (b) Western blot analysis of the presence of cleaved‐Caspase‐3 and cleaved‐PARP. (c) Western blot analysis of the expression of Beclin1, LC3B‐I, and LC3B‐II. (d) Immunofluorescence analysis of LC3B expression. LC3B was labeled green with Alexa Fluor 488‐conjugated secondary antibodies, cell nuclei were stained blue with DAPI. Scale bars: 50 μm. *p < 0.05, **p < 0.001.
Fig 2: Clinical, radiological, and genetic findings of the patient. (a) CT examination indicates bilateral renal atrophy, especially the left kidney. (b) Ultrasonic cardiogram (c) Pedigree chart of the patient's family. I1, I2, and II1 represent the father, mother, and patient, respectively, and the arrows represent the proband. Black represents DCHS1 mutation. (d) Genetic analysis shows a heterozygous DCHS1 mutation, c.8309G>A was identified in the patient, but not in her parents. (e) Sequence chromatograms of the rare (p.R2770Q) missense in silico‐predicted deleterious variants identified in this study.
Fig 3: DCHS1 DNA CNs 1 vs. ≥ 2 in subjects with UTI without VUR, UTI with VUR, and controls. Percentage and distribution of 1 CN vs. ≥ 2 CN of DCHS1 in subjects with UTI without VUR (n = 71), UTI with VUR (n = 320), and controls (n = 491). CN = DNA CN of DCHS1.
Fig 4: Efficiency of DCHS1 c.8309G>A (p.R2770Q) mutation in cultured HK‐2 cells. (a) Efficiency of DCHS1 c.8309G>A (p.R2770Q) mutation in HK‐2 cells was determined by qRT‐PCR. Control: wild‐type; DCHS1‐WT: DCHS1 was cloned to construct HK‐2 stable cell line; DCHS1‐Mut: DCHS1 (c.8309G>A, p.R2770Q) was cloned to construct HK‐2. (b) Western blot analysis of the efficiency of DCHS1‐Mut: in HK‐2 cells. (c) OD values of the three groups of cells at 24, 48, and 72 hr were measured by CCK‐8 assay. Western blot analysis of the expression of cyclin D1 and CDK4 protein. **p < 0.01.
Fig 5: DCHS1 DNA copy number (CN) in subjects with UTI without vesicoureteral reflux (VUR), UTI with VUR, and controls. DCHS1 CN distribution and percentages in subjects with UTI without VUR (n = 71), UTI with VUR (n = 320), and controls (n = 491).
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