hoechst 33 342 dye  (MedChemExpress)

Journal: Virulence

Article Title: Antibacterial efficacy and mechanism of the novel antimicrobial peptide lachnospirin-1 against Acinetobacter baumannii

doi: 10.1080/21505594.2026.2646808

Figure Lengend Snippet: Lachnospirin-1 acts on the cell membrane of AB1069. (A) Observation of the effect of lachnospirin-1 on the cell membrane under TEM. Scale: 1 µm. Red arrows indicate cell membrane rupture and content leakage; yellow arrows indicate the appearance of mesosomes. (B) Observation of PI staining fluorescence intensity under CLSM. Scale: 20 µm. (C) Quantitative analysis of PI staining. (D) Quantitative analysis of SYTOX GREEN staining. (E) Detection of membrane potential using DiSC3(5) probe. (F) Detection of membrane fluidity by Luardan staining. (G) Fluorescence labeling of FITC-lachnospirin-1 observed under CLSM, showing its localization in the cytoplasm of A. baumannii . Green fluorescence represents FITC-lachnospirin-1; red fluorescence represents the membrane stain FM4-64; blue fluorescence represents the DNA-stained Hoechst 33,342. Scale: 10 µm.

Article Snippet: FITC-lachnospirin-1 (GL Biochem Ltd., Shanghai, China) was added to the final concentration of 16 μM, and the mixture was incubated at 37°C for 1–2 h. Twenty minutes before sampling, 20 μg/mL of Hoechst 33,342 dye (MedChem Express, New Jersey, USA) was added, and 2 min before sampling, 20 μM FM4-64 fluorescent dye (MedChem Express, New Jersey, USA) was added.

Techniques: Membrane, Staining, Fluorescence, Labeling

Journal: Cell communication and signaling : CCS

Article Title: Induced mitochondrial deficit by NDUFS3 transient silencing reduces RAB7 expression and causes lysosomal dysfunction in pancreatic cancer cells.

doi: 10.1186/s12964-025-02214-y

Figure Lengend Snippet: Fig. 4 Compromised late endocytic trafficking of NDUFS3-silenced pancreatic cancer cell lines. (A-D) Relative protein abundance of RAB7, dynein, LAMP- 1, TFEB, ATP6V1G1, and RAB9 was assessed by Western blotting in YAPC and MIA PaCa-2 cells transfected with control RNA (Scr) or with two different NDUFS3 siRNAs (indicated as #1 and #2). The antibody against NDUFS3 was used to check the efficiency of RNA interference (RNAi). Quantification of differences was performed by densitometric analysis, normalizing against HSP90, and it was reported as the ratio of NDUFS3-silenced samples (#1 and #2) compared to their Scr control. (E) Late endocytic acid compartments were live stained in NDUFS3-silenced YAPC and MIA PaCa-2 cells (indicated as #1 and #2) and relative Scr controls using LysoTracker DND-99 dye (red). Nuclei were labeled with PureBluTM Hoechst 33,342 Nuclear Staining Dye (blue). White boxes indicate zoomed areas below. Scale bar: 10 μm. (F-K) Corrected total cell fluorescence (CTCF), number and size of acid compartments were determined by ImageJ software in NDUFS3-silenced YAPC and MIA PaCa-2 cells and reported as the ratio compared to their Scr control. Each measure was obtained by analyzing at least 50 cells/samples from three or more independent experiments. Values are the mean ± SEM of at least three independent experiments. *p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001

Article Snippet: Before seeding, nuclei of cells were stained with 20 nM of PureBluTM Hoechst 33,342 Nuclear Staining Dye (Bio-Rad).

Techniques: Quantitative Proteomics, Western Blot, Transfection, Control, Staining, Labeling, Fluorescence, Software