Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: A murine mesenchymal stem cell model for initiating events in osteosarcomagenesis points to CDK4/CDK6 inhibition as a therapeutic target

doi: 10.1038/s41374-021-00709-z

Figure Lengend Snippet: Murine MSCs ( A ) that have spontaneously transformed after long-term culture (B6_4, wildtype (WT) Ink4ab; B6_7, deletion (DEL) of Ink4ab; B6_10, wildtype (WT) Ink4ab) or B MSCs from Ink4ab −/− mice after transformation were treated with palbociclib for 72 h after which relative cell viability and IC 50 values were determined. Wildtype MSCs were derived from FVB mice. Data points represent the mean of three experiments performed ± standard deviation.

Article Snippet: For 2D cultures, human osteosarcoma cell lines or murine MSCs were seeded (between 3000 and 6000 cells per well) into 96-well plates and after 24 h treated with PBS or Palbociclib (dissolved in PBS, PD-0332991, Selleckchemicals, Houston, TX, USA) in concentrations ranging from 0.01 μM to 100 μM.

Techniques: Transformation Assay, Derivative Assay, Standard Deviation

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: A murine mesenchymal stem cell model for initiating events in osteosarcomagenesis points to CDK4/CDK6 inhibition as a therapeutic target

doi: 10.1038/s41374-021-00709-z

Figure Lengend Snippet: A Osteosarcoma cell lines were treated with palbociclib for 72 h after which relative cell viability and IC 50 values were determined. Cell lines MG63 and 143BHOS (green) have loss of p16, whereas other OS cell lines have intact p16 (blue). Data points represent the mean of three experiments performed ± standard deviation. B Western blot showing expression levels of Rb in osteosarcoma cell lines. GAPDH was used as a loading control. C p16 protein expression status of each osteosarcoma cell line as published was correlated with IC 50 values from the current study. n.s. not statistically significant. D CDK4 and CDK6 RNA expression levels as published elsewhere of each cell line did not correlate with IC 50 values from the current study. Each dot represents one cell line.

Article Snippet: For 2D cultures, human osteosarcoma cell lines or murine MSCs were seeded (between 3000 and 6000 cells per well) into 96-well plates and after 24 h treated with PBS or Palbociclib (dissolved in PBS, PD-0332991, Selleckchemicals, Houston, TX, USA) in concentrations ranging from 0.01 μM to 100 μM.

Techniques: Standard Deviation, Western Blot, Expressing, Control, RNA Expression

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: A murine mesenchymal stem cell model for initiating events in osteosarcomagenesis points to CDK4/CDK6 inhibition as a therapeutic target

doi: 10.1038/s41374-021-00709-z

Figure Lengend Snippet: A Hematoxylin and eosin staining of osteosarcoma MCTS treated with palbociclib for 72 h. Scalebar represents 20 µm. Inset shows SATB2 staining. B Relative cell viability and IC 50 values were determined after treatment with palbociclib. C Cleaved caspase 3 and Ki67 staining and quantification of 3D cultured MCTS of OS cell lines treated with 10 µM palbociclib (Palb) or PBS. Scale bar represents 20 µm. n.s. not statistically significant, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: For 2D cultures, human osteosarcoma cell lines or murine MSCs were seeded (between 3000 and 6000 cells per well) into 96-well plates and after 24 h treated with PBS or Palbociclib (dissolved in PBS, PD-0332991, Selleckchemicals, Houston, TX, USA) in concentrations ranging from 0.01 μM to 100 μM.

Techniques: Staining, Cell Culture

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Enzyme Replacement Therapy Can Reverse Pathogenic Cascade in Pompe Disease

doi: 10.1016/j.omtm.2020.05.026

Figure Lengend Snippet: AT-GAA Reversed the Levels of the Autophagic Markers and Alleviated the Burden of Accumulated Protein Aggregates and ER-Stress in Muscle from KO Mice Muscle biopsies were collected from age and sex-matched WT, untreated (KO), and AT-GAA-treated KO (KO-ERT) mice. (A) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with the indicated antibodies (n = 4 for each group; n = 5 for western blot with LC3 antibody). (B) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with K63-linked ubiquitin specific antibody (n = 4 for each group). Western blot with anti-GAPDH and Ponceau S staining were used as loading controls. (C) Western blot analysis of muscle lysates from WT, untreated KO (KO), and treated KO (KO-ERT) mice with anti-Grp 78 antibody (ER-stress marker; n = 4 for each group); GAPDH was used as loading control. Each data point represents an individual mouse. Statistical significance was determined by one-way ANOVA. Graphs represent mean ± SD. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: SQSTM1/p62 (#ab56416; mouse monoclonal), galectin 9 (#ab69630; rabbit polyclonal), Ubiquitin K48 (Linkage Specific) (#ab140601; rabbit monoclonal), and glyceraldehyde-3-phosphate dehydrogenase (#ab9485; rabbit polyclonal) were purchased from Abcam.

Techniques: Western Blot, Ubiquitin Proteomics, Staining, Marker, Control