passage 14 cells  (DSMZ)

Journal: International Journal of Molecular Sciences

Article Title: Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells

doi: 10.3390/ijms241311168

Figure Lengend Snippet: Phosphoproteomic analysis in HepaRG cells after treatment with MCLR and/or SB203580 (SB). HepaRG cells were treated with 50 μM SB or vehicle (V) for 1 h and then with 1 μM MCLR (M) or V for 24 h ( n = 3). Volcano plots show increased and decreased mono-phosphosites (top number). The number of increased and decreased di-phosphorylated phosphosites are shown in brackets (data points not included in volcano plots). Horizontal dotted lines indicate 1.3 log 10 p -value cut off and vertical dotted lines indicate ± 0.7 log2 fold change cutoff values used ( A – C ). Venn diagrams indicate number of shared phosphosites (percent in parentheses) between the different treatment groups ( D , E ).

Article Snippet: Undifferentiated HepaRG cells (passage 14) (cat. HPR101) were purchased from BioPredic International (Saint Grégoire, France) and cultured as per manufacturer’s protocols in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells

doi: 10.3390/ijms241311168

Figure Lengend Snippet: Phosphosite and kinase analysis of SB203580 (SB) rescue of MCLR toxicity. HepaRG cells were treated with 50 μM SB or vehicle (V) for 1 h and with 1 μM MCLR (M) or V for 24 h ( n = 3). Volcano plot shows increased and decreased mono-phosphosites (top number). The number of increased and decreased di-phosphorylated phosphosites are shown in brackets (data points not included in volcano plot). Horizontal dotted lines indicate 1.3 log 10 p -value cut off and vertical dotted lines indicate ± 0.7 log2 fold change cutoff values used ( A ). Venn diagrams indicate number of shared phosphosites (percent in parentheses) between the different treatment groups ( B , C ). Kinase perturbation analysis ( D ).

Article Snippet: Undifferentiated HepaRG cells (passage 14) (cat. HPR101) were purchased from BioPredic International (Saint Grégoire, France) and cultured as per manufacturer’s protocols in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells

doi: 10.3390/ijms241311168

Figure Lengend Snippet: Western blot analysis of PP2A, MCLR, p38, and MK2. HepaRG cells were treated with 50 μM SB (black bars) or vehicle (V) (white bars) for 1 h and with or without 1 μM MCLR (M) for 24 h. Representative blots for PP2A ( A ), MCLR ( B – D ), total and phospho-p38 ( E ), and total and phospho-MK2 ( F ). Amido black total protein stain and 2-way ANOVA tables are shown for each panel. Error bars represent mean ± standard deviation of n = 3. * p < 0.05 represent Sidak’s multiple comparison post-test comparing vehicle treatment to SB treatment within each MCLR treatment group (0 and 1 µM).

Article Snippet: Undifferentiated HepaRG cells (passage 14) (cat. HPR101) were purchased from BioPredic International (Saint Grégoire, France) and cultured as per manufacturer’s protocols in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Western Blot, Staining, Standard Deviation, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells

doi: 10.3390/ijms241311168

Figure Lengend Snippet: Western blot analysis of necroptosis and DNA damage markers. HepaRG cells were treated with 50 μM SB (black bars) or vehicle (V) (white bars) for 1 h and with or without 1 μM MCLR (M) for 24 h. Representative blots for total and phospho-MLKL ( A ), total and phospho-RIP1 ( B ), CYLD ( C ), phospho-ATR ( D ), phospho-CHK1 ( E ), and phospho-CHK2 ( F ). Amido black total protein stain and 2-way ANOVA tables are shown for each panel. Error bars represent mean ± standard deviation of n = 3. * p < 0.05 represent Sidak’s multiple comparison post-test comparing vehicle treatment to SB treatment within each MCLR treatment group (0 and 1 µM).

Article Snippet: Undifferentiated HepaRG cells (passage 14) (cat. HPR101) were purchased from BioPredic International (Saint Grégoire, France) and cultured as per manufacturer’s protocols in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Western Blot, Staining, Standard Deviation, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Involvement of the p38/MK2 Pathway in MCLR Hepatotoxicity Revealed through MAPK Pharmacological Inhibition and Phosphoproteomics in HepaRG Cells

doi: 10.3390/ijms241311168

Figure Lengend Snippet: Western blot analysis of ubiquitination and DNA damage markers. HepaRG cells were treated with 50 μM SB (black bars) or vehicle (V) (white bars) for 1 h and with or without 1 μM MCLR (M) for 24 h. Representative blots for RBX1 ( A ), SKP1 ( B ), ubiquitin ( C ), CUL4A ( D ), DDB2 ( E ), and DDB1 ( F ). Amido black total protein stain and 2-way ANOVA tables are shown for each panel. Error bars represent mean ± standard deviation of n = 3. * p < 0.05 represent Sidak’s multiple comparison post-test comparing vehicle treatment to SB treatment within each MCLR treatment group (0 and 1 µM).

Article Snippet: Undifferentiated HepaRG cells (passage 14) (cat. HPR101) were purchased from BioPredic International (Saint Grégoire, France) and cultured as per manufacturer’s protocols in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Western Blot, Ubiquitin Proteomics, Staining, Standard Deviation, Comparison