Journal: Cancer Cell International

Article Title: Quantitative HER2 profiling on circulating tumor cells using an EpCAM-independent platform in metastatic breast cancer

doi: 10.1186/s12935-025-04036-x

Figure Lengend Snippet: HER2 expression analysis in spiked SKBR3 BC cell lines. ( A ) IF images of SKBR3 cells and PBMCs recovered from HDB, either spiked with SKBR3 cells (left) or non-spiked (right). Blood was processed using the EpCAM-independent pipeline. Spike-in experiments were conducted in triplicate. The recovered cells were stained for DAPI (blue), CD45 (red), EpCAM/CK (green) and HER2 (cyan) markers. The majority of recovered SKBR3 cells were DAPI+, CD45-, EpCAM/CK+, and HER2+, whereas the majority of cells recovered from the non-spiked HDB blood were DAPI+, CD45+, EpCAM/CK-, and HER2- ( B ) Comparative bar chart showing the HER2 expression levels (high, intermediate, and low/none) established on the recovered SKBR3 cells using the EpCAM-independent pipeline.

Article Snippet: CD45 , Surface , HI30 , Alexa fluor 555 , Bioss Antibodies , bs-0522R-A555 , 1:10 , 30 , TRITC/ Rhodamine.

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Identifying membrane-bound transcriptional regulatory proteins from rare but evolutionarily conserved domain combinations

doi: 10.64898/2025.12.20.695554

Figure Lengend Snippet: PREB without its transmembrane domain undergoes nuclear translocation and binds to the butyrophilin locus. ( a ) Predicted structure of PREB generated by AlphaFold, highlighting the WD repeat domains, the transmembrane domain, and the site of an engineered C-terminal truncation. ( b ) Schematic of the two constructs created to assay the mouse full-length PREB protein and PREBΔTM, lacking the C-terminal transmembrane domain. The N-terminal biotinylation site is indicated. ( c ) Western blot results of two biological replicates of ES cells expressing PREB and PREBΔTM. Two bands for PREB were detected by Streptavidin-HRP, suggesting C-terminal processing of a full-length PREB when expressed in mouse ES cells. ( d ) Immunofluorescence confocal microscopy images of PREB and PREBΔTM following Dox induction. PREB localizes predominantly to a perinuclear region, forming a ring-like shape. PREBΔTM displays diffused nuclear localization. Rightmost panels show zoomed images corresponding to the boxed regions. ( e ) bioChIP-seq profiles for PREB, PREBΔTM, BirA-only Strep-pulldown, and IgG ChIP-seq control samples. PREBΔTM samples show substantially stronger DNA-binding activity than full-length PREB. ( f ) Representative locus-specific binding profiles of PREB and PREBΔTM at the Btn1a1 (butyrophilin subfamily 1 member A1) gene. PREBΔTM shows markedly higher peak intensities, consistent with enhanced nuclear localization in the absence of the transmembrane domain.

Article Snippet: Cells were then incubated overnight at 4°C with Streptavidin conjugated to Alexa Fluor 555 (Thermo Fisher Scientific, S21381) in PBS-CM containing 0.1% BSA and 10% horse serum.

Techniques: Translocation Assay, Generated, Construct, Western Blot, Expressing, Immunofluorescence, Confocal Microscopy, ChIP-sequencing, Control, Binding Assay, Activity Assay