Journal: International Journal of Molecular Sciences

Article Title: Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown

doi: 10.3390/ijms222111738

Figure Lengend Snippet: Mitophagy evaluation on 3T3-L1 adipocytes exposed to CDCA in the absence or presence of TGR5. Fluorescence microscopy representative images of the various experimental conditions, when observed under white light or evaluating fluorescent signal of the two probes, MtPhagy and Lysotracker Green. A fourth panel indicates the fluorescence signals’ colocalization where it happens, where it is represented in yellow. Scale bars in microscopy photographs represent a distance of 22 µm. Microscopy figures contrast settings were all maxed solely to gain visual clarity and long after image analysis. Bars represent means ± SEM of at least 185 individually quantified adipocytes. * indicates a statistically significant difference vs. Control ( p < 0.05); & indicates a statistically significant difference vs. Cas9 ( p < 0.05).

Article Snippet: Finally, cells were washed twice with HEPES-buffered Hanks and imaged by confocal fluorescence microscopy (Maker and model: Carl Zeiss LSM 710; Type, magnification and numerical aperture of the objective lens: Plan Apochromat 63x/1.4; Temperature: controlled room temperature of 20 °C; Medium: Hanks Balanced Salt Solution, (Sigma-Aldrich); Fluorochromes: Lysotracker Green and Mtphagy dye, both from Dojindo EU, Munich, Germany; Camera maker and model: Not applicable; Acquisition software: Zen Black, Carl Zeiss, Jena, Germany).

Techniques: Fluorescence, Microscopy, Control