Journal: Aging (Albany NY)

Article Title: SAMP-Score: a morphology-based machine learning classification method for screening pro-senescence compounds in p16 positive cancer cells

doi: 10.18632/aging.206333

Figure Lengend Snippet: Validation of QM0005928 (QM5928). ( A ) QM0005928 chemical structure. ( B ) Heatmap representing high content analysis feature (HCA; y-axis) profiles of a dose response of QM5928 (x-axis). ( C ) Line plot showing change in SAMP score with increasing doses of QM5928. ( D ) Raw cell counts of increasing doses of QM5928 (Red = NonSen and Blue = Sen SAMP-Score Classification). ( E , F ) Z-scores for cell count and cell area increasing doses of QM5928. (Red = NonSen and Blue = Sen SAMP-Score Classification). ( G ) Western plot showing p16 expression in MB-468, BT549 and HeLa Sen-Mark+ cancer lines. ( H , I ) Cell counts for MB-468, BT549 and HeLa Sen-Mark+ cancer lines in response to QM5928. N = 3. Scale bar = 100 μm. ( J ) Heatmap representing high content analysis feature (HCA; y-axis) profiles of HeLa screen treatments that phenocopy QM5928 by hierarchical clustering. ( K ) KEGG pathways analysis of HeLa screen treatments that phenocopy QM5928 by hierarchical clustering. The data for MB-468s in and are derived from the same experimental dataset, presented in different formats to emphasise distinct aspects of the findings.

Article Snippet: HeLa (CRUK), MDA-MB-468 (ATCC, HTB-132; referred to as MB-468s) BT549 (ATCC, HTB-122) and MDA-MB-231 (ATCC, HTB-26; referred to as MB-231s) cells were cultured in high-glucose DMEM (Life Technologies, UK).

Techniques: Biomarker Discovery, High Content Screening, Cell Characterization, Western Blot, Expressing, Derivative Assay

Journal: Aging (Albany NY)

Article Title: SAMP-Score: a morphology-based machine learning classification method for screening pro-senescence compounds in p16 positive cancer cells

doi: 10.18632/aging.206333

Figure Lengend Snippet: Interplay between p16 localisation and QM5928. ( A ) Immunofluorescence staining of MB-468s treated with either DMSO or QM5928 for DAPI, p16 (Santa Cruz - FITC), p16 (Protein Tech - Cy3) and Cell Mask. Secondary only controls (2nd) did not receive primary p16 antibodies. ( B ) Digital zoom of white boxed areas in A, for p16 (Protein Tech - Cy3). Scale bars = 100 μm. ( C , D ) Scaled probability density estimate quantitation of nuclear/cytoplasmic intensity ratios for MB-468s stained with p16 in the FITC and Cy3 channels. A higher value indicates increased nuclear staining. Area under the curve is equal to 100% of the population. N = 3. ( E , F ) Cell counts for MB-468 (p16 positive - blue) and MB-231 (p16-null - red) cancer lines in response to QM5928 at day 6 or day 10. N = 3. The data for MB-468s in and are derived from the same experimental dataset, presented in different formats to emphasise distinct aspects of the findings.

Article Snippet: HeLa (CRUK), MDA-MB-468 (ATCC, HTB-132; referred to as MB-468s) BT549 (ATCC, HTB-122) and MDA-MB-231 (ATCC, HTB-26; referred to as MB-231s) cells were cultured in high-glucose DMEM (Life Technologies, UK).

Techniques: Immunofluorescence, Staining, Quantitation Assay, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Rice Peroxygenase-9 Negatively Regulates Production of Reactive Oxygen Species and Increases Cellular Resistance to Abiotic Stress

doi: 10.3390/ijms26146918

Figure Lengend Snippet: Accumulation of ROS and cell death level in rice seedlings from WT and ox lines under drought and salt stress. ( A ) DAB staining, ( B ) NBT staining, and ( C ) Evan’s blue staining. Three-week-old seedlings grown in soil were subjected to stressors, as follows: For drought stress, seedlings were taken out and dried in the air at room temperature for 4 h (DAB and NBT staining) or 6 h (Evan’s blue staining); for salt stress, 200 mM NaCl was added directly to the water reservoir containing the rice seedling pots for 24 h (DAB and NBT staining) or 36 h (Evan’s blue staining). All the treated rice seedlings were re-subjected to the normal water condition for 12 h to recover before staining with 0.2% DAB ( w/v ) solution (24 h), 0.1% ( w/v ) NBT solution (24 h), or 0.25% Evan’s blue ( w/v ) solution (30 min), followed by boiling in 95% ethanol for 4 h and observed under a light microscope with 10× magnification. Non-treated seedlings were used as the control. White bar = 1 cm and black bar = 1 mm. The stained area was converted from three separated microscopic photos using ImageJ Version 1.45p. ( D ) Malondialdehyde (MDA) content in the leaves of WT, ox2-1, and ox2-2 under drought and salt stress. Error bars denote ± SD of three replicates. The p -values were calculated using the Student’s t -test (** p < 0.05 and *** p < 0.01).

Article Snippet: For MDA content quantification, third leaves of the WT and stress-treated seedlings were collected, and MDA content was extracted and measured by the Malondialdehyde (MDA) Colorimetric Assay Kit (Cat. #E-BC-K025-S, Elabscience, Houston, TX, USA).

Techniques: Staining, Light Microscopy, Control