western blot wb  (Proteintech)

Journal: iScience

Article Title: ZBED6–IGF2–PIK3C3 autophagy axis drives ccRCC progression: A multi-omics integration study

doi: 10.1016/j.isci.2026.114952

Figure Lengend Snippet: Preliminary verification of the regulatory relationship between ZBED6 and PIK3C3 (A) Scatterplot shows the correlation between ZBED6 and PIK3C3 in ArrayExpress (left) and ICGC-RECU (right). (B) Bar graph illustrates the expression levels of ZBED6 and PIK3C3 across different cell lines. (C) Western blot (WB) analysis validates differential protein expression of ZBED6 and PIK3C3 in renal tubular epithelial cell lines and clear cell renal cell carcinoma (ccRCC) cell lines. (D) WB analysis demonstrates changes in ZBED6 and PIK3C3 expression in the 769-P cell line following ZBED6 knockdown and overexpression. (E) Subcellular localization of ZBED6 in cells based on the Human Protein Atlas (HPA) database. (F) Immunohistochemical (IHC) results from HPA show PIK3C3 expression in ccRCC tumor tissues versus normal renal tissues. (G) Altered expression of autophagy markers after ZBED6 overexpression. ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, ∗∗∗ indicates p < 0.001, and ∗∗∗∗ indicates p < 0.0001.

Article Snippet: Preliminary verification of the regulatory relationship between ZBED6 and PIK3C3 (A) Scatterplot shows the correlation between ZBED6 and PIK3C3 in ArrayExpress (left) and ICGC-RECU (right). (B) Bar graph illustrates the expression levels of ZBED6 and PIK3C3 across different cell lines. (C) Western blot (WB) analysis validates differential protein expression of ZBED6 and PIK3C3 in renal tubular epithelial cell lines and clear cell renal cell carcinoma (ccRCC) cell lines. (D) WB analysis demonstrates changes in ZBED6 and PIK3C3 expression in the 769-P cell line following ZBED6 knockdown and overexpression. (E) Subcellular localization of ZBED6 in cells based on the Human Protein Atlas (HPA) database. (F) Immunohistochemical (IHC) results from HPA show PIK3C3 expression in ccRCC tumor tissues versus normal renal tissues. (G) Altered expression of autophagy markers after ZBED6 overexpression.

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Immunohistochemical staining

Journal: bioRxiv

Article Title: Fine-tuning STEAP1 protein expression and purification to preserve its conformation and function

doi: 10.64898/2026.02.16.706263

Figure Lengend Snippet: A) Diagram of STEAP1 constructs. mEGFP: monomeric enhanced green fluorescent protein. B) Cell viability and VCD (Viable Cell Density) of cells expressing STEAP1 at different culturing timepoints. C) Western blot analysis of STEAP1 expression levels in relation to varying culturing durations. The asterisk denotes bands of STEAP1 positioned at the expected molecular weight. Arrows indicate bands of STEAP1 corresponding to the size of dimers and trimers. D) Measurement of STEAP1 expression levels in cells treated with heme additives (0.5 mM 5-Aminolevulinic acid HCl, 1 μM Hemin-Cl, and 5 μM Fe(III)Cl) or not via western blotting. An equal number of cells were used to prepare samples for each lane. E) FSEC analysis of the oligomeric status of STEAP1-mEGFP in cells treated with heme additives compared to those without additives. F) Cryo-EM structure of STEAP1 homotrimers (PDB: 8UCD), corresponding to the trimeric peak in . The heme moiety is shown in red, lipid in pink, and bound FAD in magenta.

Article Snippet: Following SDS-PAGE, proteins were transferred to nitrocellulose membranes, which were blocked using Blocking Buffer for Fluorescent Western Blotting (cat. no. MB-070, Rockland Immunochemicals) and incubated with monoclonal anti-FLAG antibodies (cat. no. F1804, Sigma).

Techniques: Construct, Expressing, Western Blot, Molecular Weight, Cryo-EM Sample Prep