Journal: Microsystems & Nanoengineering

Article Title: Integration of multiple flexible electrodes for real-time detection of barrier formation with spatial resolution in a gut-on-chip system

doi: 10.1038/s41378-023-00640-x

Figure Lengend Snippet: a Impedance magnitude and b phase recorded for 12 days of Caco-2 cell culture inside HuMiX. c Immunofluorescence staining of the epithelial Caco-2 barrier grown in the HuMiX for 12 days. Cell nuclei were stained with DAPI (blue) and the tight junction protein occludin was stained with Alexa Fluor 488 (green). The measurements were taken at electrode position 2 (~29 mm from the inlet of the device)

Article Snippet: The cells were labeled with occludin antibody diluted in 4% BSA for 3 h at room temperature, and nuclei were stained overnight with Fluoroshield TM with DAPI (F6057, Merck Milipore, Hoeilaart, Belgium).

Techniques: Cell Culture, Immunofluorescence, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Alternative splicing regulates PACC1 function and promotes acidosis-induced cytotoxicity

doi: 10.3389/fcell.2025.1754079

Figure Lengend Snippet: Expression of PACC1 splice isoforms in vitro . (A) Schematic representation of the human PACC1-V2 structure (AlphaFold code: AF- Q9H813 -F1-v4), highlighting the position of the amino acid sequence encoded by the alternative exon 2 present in PACC1-V1. (B,C) Western blot analysis of PACC1 isoforms in total lysates from HEK-293 PACC1 KO cells transiently transfected with either PACC1-V1 or PACC1-V2. Isoforms were detected using an anti-HA antibody under denaturing (with β-mercaptoethanol, βME) and non-denaturing (without βME) conditions. GAPDH was used as a loading control (bottom images). Original Western blot images. (C) Densitometric analysis of Western blot bands showing quantification of monomeric (left panel) and dimeric (right panel) forms of PACC1. **, p < 0.01 by Student’s t-test (n = 4). (D,E) Immunofluorescence detection of PACC1 splice isoforms in transfected PACC1-KO cells. (D) Quantification of transfection efficiency (top) and fluorescence intensity (bottom) for PACC1-V1 and PACC1-V2. **, p < 0.01 by Student’s t-test (n = 6 independent transfections, each with 5 technical replicates). (E) Representative confocal images showing anti-HA immunostaining of PACC1 (magenta); nuclei were counterstained with DAPI (cyan). Scale bar: 5 μm. Intensity profiles of PACC1 staining along the dashed white lines in the images are shown in the adjacent graphs.

Article Snippet: After three additional PBS washes, chambers were removed, and slides were mounted using Fluoroshield mounting medium with DAPI (Merck, F6057) to counterstain nuclei.

Techniques: Expressing, In Vitro, Sequencing, Western Blot, Transfection, Control, Immunofluorescence, Fluorescence, Immunostaining, Staining