Journal: bioRxiv

Article Title: NSD2 promotes cell durotaxis and drives the transition from PKD to tubulocystic renal cell carcinoma through integrin/FAK/AKT signaling

doi: 10.1101/2024.10.21.619559

Figure Lengend Snippet: (A) Normalized read density of H3K36me2 CUT&Tag signals of Ksp Cre ; Nsd2 OE/+ and Nsd2 OE/+ mice at 12 weeks from 3 kb upstream of the TSS to 3 kb downstream of the TES. (B) KEGG pathway analysis of gene expression changes in CUT&Tag data. (C) Venn diagram showing the number of genes harboring H3K36me2 binding and the differential expression genes determined by RNA-seq data. (D) The KEGG pathway analysis of the overlapping genes in (C). (E) RT-qPCR analysis of integrin family mRNA levels of KM and KMN mice (n = 3 per group). Experiments were repeated at least three times, with similar results. (F) RT-qPCR analysis of ITGA4 and ITGA11 mRNA levels of EV and Nsd2-OE cells (Caki-1 and 293T cells derived, n = 3 per group). Experiments were repeated at least three times, with similar results. (G) RT-qPCR analysis of ITGA4 and ITGA11 mRNA levels of Ctrl and Nsd2-KO cells (Caki-1 and 293T cells derived, n = 3 per group). Experiments were repeated at least three times, with similar results. (H) IHC staining and quantification of ITGA4 and ITGA11 expressions in KM and KMN mice at 12 weeks (n = 5 per group). Experiments were repeated at least three times, with similar results, and representative images are shown. Scale bars: 50 μm. (I) Snapshot of H3K36me2 CUT&Tag signals at the Itga4 and Itga11 gene in Nsd2 OE/+ and Ksp Cre ; Nsd2 OE/+ mice. (J) ChIP-qPCR of Itga4 and Itga11 in Nsd2 OE/+ and Ksp Cre ; Nsd2 OE/+ mice. The location of the ChIP primer pairs used is at the promoter area. Experiments were repeated at least three times, with similar results. (K) The Pearson product-moment pair-wise gene correlation analysis between NSD2 and ITGA4 , ITGA11 with TCGA database.

Article Snippet: Next, samples were blocked with 5% BSA at room temperature for 1 h. Then, samples were incubated with primary antibodies at 4 °C for 12-16 h. The primary antibodies used were NSD2 (Abcam, ab75359), H3K36me2 (Abcam, ab9049), AMACR (Abcam, ab140798), Vimentin (Cell Signaling Technology, 5741), Ki67 (Abcam, ab15580), CK19 (Abcam, ab15463), Collagen I (Cell Signaling Technology, 72026), ITGA11 (Abcam, ab198826), ITGA4 (Cell Signaling Technology, 8440), p-FAK (Cell Signaling Technology, 3283), p-AKT (Cell Signaling Technology, 4060), Collagen V (Abcam, ab7046).

Techniques: Gene Expression, Binding Assay, Quantitative Proteomics, RNA Sequencing, Quantitative RT-PCR, Derivative Assay, Immunohistochemistry, ChIP-qPCR

Journal: bioRxiv

Article Title: NSD2 promotes cell durotaxis and drives the transition from PKD to tubulocystic renal cell carcinoma through integrin/FAK/AKT signaling

doi: 10.1101/2024.10.21.619559

Figure Lengend Snippet: (A) Scheme of treatment given for each gavage. (B) Kidney images of KMN mice treated with vehicle or Firategrast for 21 days. Quantification of kidney volume (mm 3 ) is on the right (n = 6 per group). (C) Representative hematoxylin and eosin images of kidney tissue from KMN mice treated with vehicle or Firategrast for 21 days. Scale bars: 100 μm. Quantification of cystic index (%) and number of cysts per field is shown at the bottom (n = 6 per group). (D) IF analysis of Collagen I expression and IHC staining of p-FAK, ITGA4, and ITGA11 in KMN mice treated with vehicle or Firategrast for 21 days. Scale bars: 50 μm. Quantification of Collagen I, p-FAK, ITGA4 and ITGA11 positive area is shown at the bottom (n = 6 per group). (E) IHC staining and quantification of CK19, AMACR, Vimentin positive area, and Ki67 positive cells per field from indicated mice (n = 6 per group). Scale bars: 50 μm. (F) CCK8 assay of EV and Nsd2-OE cells transfected with the indicated plasmid, respectively. Experiments were repeated at least three times, with similar results. (G) CCK8 assay of EV and Nsd2-OE cells treated with MK2206 or vehicle, respectively. Experiments were repeated at least three times, with similar results.

Article Snippet: Next, samples were blocked with 5% BSA at room temperature for 1 h. Then, samples were incubated with primary antibodies at 4 °C for 12-16 h. The primary antibodies used were NSD2 (Abcam, ab75359), H3K36me2 (Abcam, ab9049), AMACR (Abcam, ab140798), Vimentin (Cell Signaling Technology, 5741), Ki67 (Abcam, ab15580), CK19 (Abcam, ab15463), Collagen I (Cell Signaling Technology, 72026), ITGA11 (Abcam, ab198826), ITGA4 (Cell Signaling Technology, 8440), p-FAK (Cell Signaling Technology, 3283), p-AKT (Cell Signaling Technology, 4060), Collagen V (Abcam, ab7046).

Techniques: Expressing, Immunohistochemistry, CCK-8 Assay, Transfection, Plasmid Preparation

Journal: eLife

Article Title: Proteomic landscape of tunneling nanotubes reveals CD9 and CD81 tetraspanins as key regulators

doi: 10.7554/eLife.99172

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-ITGB1, ITGB4, ITGA4 , Cell Signaling , #4749 , WB (1/1000).

Techniques: Transfection, Construct, Expressing, Plasmid Preparation, Marker, Sequencing, Purification, Transduction, Control, Software, Staining