Journal: International journal of molecular sciences

Article Title: Porcine Kidney Organoids Derived from Naïve-like Embryonic Stem Cells.

doi: 10.3390/ijms25010682

Figure Lengend Snippet: Figure 5. Gene expression in 3D porcine kidney organoids. (A) Expression of renal progenitor cell markers in the organoids on day 21 and fetal porcine kidney on day 45. Renal progenitor cells markers (EYA1, SIX1) were measured by immunofluorescence test (EYA1 was localized in the cytoplasm, SIX1 was localized in the nucleus). (B) Expression of mature renal cell markers in the organoids on day 21 and fetal porcine kidney on day 45. Mature nephron components markers (PAX2, E-CAD, PODO) were measured by immunofluorescence test and immunohistochemistry test (the circular dotted lines are tubule-like structures; PAX2 was localized in the nucleus; and PODO and ECAD were localized to the cell membrane). (C) Whole-mount co-staining of the organoids for WT1 and CD31 on day 21 (WT1 was localized in the nucleus and CD31 was localized to the cell membrane) (scale bar: 50 µm).

Article Snippet: The primary antibodies included the following: EYA1 (Abcam), SIX1 (Proteintech), PAX2 (Santa Cruz Biotechnology), WT1 (Abcam), E-CAD (Abcam), PODO (Absin), CD31 (Santa Cruz Biotechnology), AQP1 (Proteintech), GLUT1 (proteintech), and KI67 (Abcam); the secondary antibodies included goat anti-rabbit IgG Alexa Fluor 546 (ABclonal), goat anti-rabbit IgG Alexa Fluor 488 (Thermo Fisher Scientific), goat anti-mouse IgG Alexa Fluor 546 (Thermo Fisher Scientific), goat anti-mouse IgG Alexa Fluor 488 (Thermo Fisher Scientific), goat anti-rabbit IgG H&L(HRP) (Abcam), and goat anti-mouse IgG(H + L) (KeyGEN BioTECH, Nanjing, China).

Techniques: Gene Expression, Expressing, Immunohistochemistry, Membrane, Staining

Journal: Molecular Metabolism

Article Title: P2Y2R contributes to the development of diabetic nephropathy by inhibiting autophagy response

doi: 10.1016/j.molmet.2020.101089

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Wilms Tumor-1 (WT1) , Boster Biological Technology , #M00199-1 , Rabbit , 1 μg/ml (IHC).

Techniques:

Journal: Molecular Metabolism

Article Title: P2Y2R contributes to the development of diabetic nephropathy by inhibiting autophagy response

doi: 10.1016/j.molmet.2020.101089

Figure Lengend Snippet: P2Y2R deficiency protects against podocyte loss and glomerular injury in DN. (A) Relative mRNA levels of Nphs1 (nephrin) and Nphs2 (podocin) were determined by real-time PCR analysis (n = 3–5). (B) Localisation of WT1, a podocyte marker, was analysed by immunohistochemistry, and representative images are shown. The number of stained podocytes per glomeruli was counted by using ImageJ software (n = 3). (C) Kidney sections were stained with PAS staining and glomerular morphological changes were scored as described in the method section (n = 3). Data are presented as mean ± SEM. One-way ANOVA was used for statistical analysis followed by Bonferroni's multiple comparisons test. ∗∗∗p < 0.001 vs WT control mice; and ### p < 0.001 vs WT DN mice. Scale bar, 50 μm.

Article Snippet: Wilms Tumor-1 (WT1) , Boster Biological Technology , #M00199-1 , Rabbit , 1 μg/ml (IHC).

Techniques: Real-time Polymerase Chain Reaction, Marker, Immunohistochemistry, Staining, Software, Control

Journal: Scientific Reports

Article Title: Identification of m6A methyltransferase-related WTAP and ZC3H13 predicts immune infiltrates in glioblastoma

doi: 10.1038/s41598-025-88671-4

Figure Lengend Snippet: Relative expression of genes expression in GBM tissues. Expression levels of WTAP and ZC3H13 genes were compared with those of the control group on GBM tissues, 10 GBM and adjacent tissues, respectively ( A ). The expression levels of siRNA WTAP and overexpression ZC3H13 genes ( B and C ). The expression levels of CD27, CD70, CD80, CD86, ICOS, CTLA4, and LAG3 in siRNA WTAP and overexpression ZC3H13 were demonstrated in Fig. 9D and F, respectively. The result were determined by qRT-PCR. The data are mean ± standard deviation of six replicate experiments. * P < 0.05, **P < 0.01 and ***P < 0.001 compared with the corresponding control values.

Article Snippet: After blocking with 5% nonfat milk, the membranes were immunoblotted with the primary antibodies: WTAP (A04296-2, Boster Biotech, Wuhan, China), CD27 (A01148-2, Boster Biotech), CD70 (A02853-2, Boster Biotech), CD80 (A00196-3, Boster Biotech), CD86(BM4121, Boster Biotech), ICOS (A00291-2, Boster Biotech), CTLA4 (A00020-1, Boster Biotech), LAG3 (M02869-2, Boster Biotech), Beta-actin (BM3873, Boster Biotech).

Techniques: Expressing, Control, Over Expression, Quantitative RT-PCR, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: APT1-derived depalmitoylation of CD36 alleviates diabetes-induced lipotoxicity in podocytes

doi: 10.7150/ijbs.109220

Figure Lengend Snippet: Construction of db/db model to evaluate changes in renal pathology lipid metabolism. Two groups of twenty-week-old mice were analyzed: db/m (n=6) and db/db (n=6). A-B. Representative PAS staining of glomeruli in each group, and the semiquantitative analysis of mesangial area ratio. (**<0.01, scale bar: 50 μm). C-G. Biochemical examination: (C) UACR, (D) serum creatinine, (E) blood glucose, (F) serum cholesterol, (G) serum triglycerides. (**<0.01). H. Representative transmission electron microscopy (TEM) images of podocyte foot processes and glomerular basement membrane (GBM) in each group. (**<0.01, scale bar: 10 μm). I-J. Representative immunofluorescence double staining of BODIPY and WT1 in the glomeruli of db/m and db/db mice, and semiquantitative analysis. (**<0.01, scale bar:50μm). K-L. Oil Red O staining of lipid droplets (LDs) in the glomeruli of db/m and db/db mice, and semiquantitative analysis. (**<0.01, scale bar: 40 μm).

Article Snippet: Frozen kidney sections were fixed, blocked, and then stained with WT1 antibody (BM4216, Boster, China) for 1 h at 37°C, subsequently incubated in the presence of secondary antibody and BODIPY dye, and finally covered with DAPI.

Techniques: Staining, Transmission Assay, Electron Microscopy, Membrane, Immunofluorescence, Double Staining