phenol red free cell culture medium  (PromoCell)

Journal: Apoptosis

Article Title: CD24 flags anastasis in melanoma cells

doi: 10.1007/s10495-024-01990-1

Figure Lengend Snippet: Quantification of secreted pigment in 3D culture supernatants. Assessment of pigment synthesizing potential of Susp and Adh subpopulations of B16-F10 cells by spectrophotometric measurement of the optical density of the culture supernatants from the 3D colony formation assay where cells were cultivated in dye-free complete RPMI medium. Inset shows images of the culture supernatant from the media control, Adh, and Susp 3D culture, respectively

Article Snippet: For this purpose, 3D soft agarose culture assays were set up using phenol red-free RPMI (ThermoFischer Scientific, 11,835,030) complete cell culture medium.

Techniques: Colony Assay, Control

Journal: Nature Chemical Biology

Article Title: A general method for the development of multicolor biosensors with large dynamic ranges

doi: 10.1038/s41589-023-01350-1

Figure Lengend Snippet: a , Schematic representation of ChemoG–ATP; NTD, N-terminal domain. b , Fluorescence intensity emission spectra of SiR-labeled ChemoG–ATP at different ATP concentrations. Means of three technical replicates are shown; [ATP], ATP concentration. c , ATP titration curves of ChemoX–ATP SiR sensors. Data are shown as the means ± s.d. of the FRET/eGFP ratio changes (Δ R / R 0 ; n = 3 technical replicates). The intracellular ATP concentration range is indicated with a gray box. Δ R / R 0 and C 50 values are summarized in Supplementary Table . d , Confocal images of HeLa Kyoto cells expressing ChemoG–ATP labeled with SiR. Shown are the eGFP channel, the FRET channel and the ratio image of both channels (FRET/eGFP) in pseudocolor (LUT = mpl-viridis). Cells were treated at t = 5 min with 10 mM 2DG. At t = 20 min, 20 mM glucose (Glc) was added to the cells until the end of the experiment ( t = 35 min, 2DG + Glc); scale bars, 25 µm. e , Time course measurement of ChemoG–ATP SiR fluorescence intensity in HeLa Kyoto cells. Shown are the eGFP and FRET channels (left) and FRET/eGFP ratio (right) normalized to 1 at t = 0 min. Cells were treated with 10 mM 2DG and subsequently with 20 mM glucose at time points indicated with arrows. Experiments are as explained in d ; n = 59 cells from three biological replicates. Represented are the means (solid lines) and traces of the individual cells (dim lines). f , Time course measurement of ChemoB–ATP SiR ( n = 58 cells), ChemoG–ATP SiR ( n = 63 cells), ChemoR–ATP SiR ( n = 52 cells) and ATeam 1.03 ( n = 59 cells) fluorescence intensity in HeLa Kyoto cells. The FRET/FP ratio after treatment with 10 mM 2DG is shown. Ratios are normalized to 1 at t = 0 min. Addition of 2DG is indicated with an arrow. Represented are the means (line) and single-cell traces (dim lines) from three biological replicates.

Article Snippet: Media were exchanged 1 h before the start of the time course with the following changes: HeLa Kyoto cells, ATP sensors, phenol red-free cell culture medium without glucose (Gibco), U-2 OS cells, NAD + sensors, phenol red-free 1× HBSS with calcium and magnesium (Corning) and HeLa Kyoto cells, calcium sensors, phenol red-free cell culture medium (Gibco).

Techniques: Fluorescence, Labeling, Concentration Assay, Titration, Expressing

Journal: Nature Chemical Biology

Article Title: A general method for the development of multicolor biosensors with large dynamic ranges

doi: 10.1038/s41589-023-01350-1

Figure Lengend Snippet: a , Schematic representation of ChemoL–NAD. b , Luminescent intensity (LI) emission spectra of CPY-labeled ChemoL–NAD at different NAD + concentrations. Means of three technical replicates are shown. c , NAD + titration curve of ChemoL–NAD CPY . Shown are the mean BRET–FRET CPY /eGFP luminescence ratios ± s.d of three technical replicates. d , ChemoL–NAD CPY BRET–FRET/eGFP ratios in U-2 OS cells after treatment for 24 h with DMSO (Ctrl), 1 mM NR, 100 nM FK866 or 100 nM FK866 and 1 mM NR. Represented are the means ± s.d. and single-well ratios (circles; n = 18 wells per condition from three biological replicates). P values are given based on unpaired two-tailed t -tests with Welch’s correction; ** P = 0.006; **** P < 0.0001. e , Time course measurement of ChemoL–NAD CPY expressed in U-2 OS cells. Represented are the means of the BRET–FRET/eGFP ratios (line) ± s.d. (shaded areas) normalized to 1 at t = 0 min. Cells were untreated (+medium) or treated (+MNNG) with MNNG at t = 5 min indicated with an arrow ( n = 3 wells from one representative biological replicate; two additional biological replicates can be found in Supplementary Fig. ). f , Time course measurement of ChemoL–ATP CPY expressed in HeLa Kyoto cells. Represented are the mean BRET–FRET/eGFP ratios (line) ± s.d. (shaded areas) normalized to 1 at t = 0 min. Cells were untreated (medium), treated with 2DG at t = 5 min (red and orange) and additionally treated with glucose at t = 25 min (orange). Addition of medium, 2DG and glucose is indicated with an arrow ( n = 3 wells from one representative biological replicate; two additional biological replicates can be found in Supplementary Fig. ). g , Time course measurement of ChemoL–CaM CPY expressed in HeLa Kyoto cells. Shown are the mean BRET–FRET/eGFP ratios (line) ± s.d. (shaded areas) normalized to 1 at t = 0 min. Cells were untreated or treated with histamine or ionomycin at t = 2 min ( n = 3 wells from one representative biological replicate; two additional biological replicates can be found in Supplementary Fig. ). Addition of drugs is indicated with an arrow.

Article Snippet: Media were exchanged 1 h before the start of the time course with the following changes: HeLa Kyoto cells, ATP sensors, phenol red-free cell culture medium without glucose (Gibco), U-2 OS cells, NAD + sensors, phenol red-free 1× HBSS with calcium and magnesium (Corning) and HeLa Kyoto cells, calcium sensors, phenol red-free cell culture medium (Gibco).

Techniques: Labeling, Titration, Two Tailed Test