Journal: Journal of Nucleic Acids

Article Title: DNA Ligase IV Prevents Replication Fork Stalling and Promotes Cellular Proliferation in Triple Negative Breast Cancer

doi: 10.1155/2019/9170341

Figure Lengend Snippet: Lig4 depletion does not increase Chk1 or Chk2 phosphorylation or dephosphorylation. pChk1 (Ser317 or Ser345), pChk2 (Thr68), total Chk1, or total Chk2 were assessed via western blot. The experiment was repeated three times. (a, c) Lig4 depleted or scrambled control treated (NT) cells were treated with 10 mM hydroxyurea or PBS for 1 or 24 hr. (b, d) Lig4 depleted or NT cells were treated with 10 mM HU or PBS for 1 hr and released for indicated times.

Article Snippet: The following primary antibodies were used: Lig4, DNA-PKcs and pDNA-PKcs S2056 (Abcam, Cambridge, UK), β -actin and Vinculin (Santa Cruz Biotechnology, Santa Cruz, CA) and GAPDH, cleaved caspase 3, caspase 9, β -tubulin, pChk1-S317, pChk1-S345, total Chk1, pChk2-T68, or total Chk2 (Cell Signaling Technology, Danvers, MA).

Techniques: De-Phosphorylation Assay, Western Blot

Journal: iScience

Article Title: Lipopolysaccharide induces placental mitochondrial dysfunction in murine and human systems by reducing MNRR1 levels via a TLR4-independent pathway

doi: 10.1016/j.isci.2022.105342

Figure Lengend Snippet: ROS generated by NOX2 activates ATM kinase in LPS-treated placental cells (A–C) Left , Equal amounts of HTR cells treated with control (water) or LPS were separated on an SDS-PAGE gel and probed for NOX2. Tubulin was probed as a loading control. Right , NOX2 levels relative to tubulin are shown (n = 4 biological replicates). (B) Left , Equal amounts of HTR cells were treated for 24 h with control (water) or LPS and, for second blot, 25 μM NOX2 inhibitor (using DMSO in control); lysates were separated on an SDS-PAGE gel and probed for MNRR1. Actin was probed as a loading control. Right , Relative MNRR1 levels are shown for each lane (n = 4 biological replicates). (C) HTR cells were treated with control (water) or LPS for the times shown, and ROS levels were measured as in Figure 1 C. Total ROS, black; mitochondrial ROS, red; total ROS with ATM inhibitor, gray (n = 2 biological replicates). (D) Equal amounts of HTR cells were treated with control (water) or H 2 O 2 for 16 h and lysates separated on an SDS-PAGE gel and probed for phospho-CHK2, total CHK2, and ATM kinase. Actin was probed as a loading control. (E) Left, Equal amounts of HTR cells treated with control (water) or LPS with either Vehicle (DMSO) or 100 μM N-acetyl cysteine for 24 h were separated on an SDS-PAGE gel and probed for YME1L1. Actin was probed as a loading control. Right , Relative YME1L1 levels are shown for each condition (n = 4 biological replicates). (F) TNFα and PTGS2 transcript levels relative to Actin were measured in HTR cells treated with Control (water), LPS, or LPS +20 μM MitoTempo (n = 4 biological replicates). (G) TNFα and PTGS2 relative transcript levels in HTR cells treated with Control (DMSO), LPS (LPS + DMSO) or LPS +25 μM NOX2 inhibitor (n = 4 biological replicates). (H) Equal amounts of YME1L1 −/− cells overexpressing WT or various mutants of YME1L1 were treated as control (water) or with LPS (1 μg/mL). Lysates were separated on an SDS-PAGE gel and probed for MNRR1, STARD7, and YME1L1. Actin was probed as a loading control.

Article Snippet: α-total CHK2 , Cell Signaling Technology , Cat # 6334; RRID: AB_11178526.

Techniques: Generated, Control, SDS Page

Journal: iScience

Article Title: Lipopolysaccharide induces placental mitochondrial dysfunction in murine and human systems by reducing MNRR1 levels via a TLR4-independent pathway

doi: 10.1016/j.isci.2022.105342

Figure Lengend Snippet:

Article Snippet: α-total CHK2 , Cell Signaling Technology , Cat # 6334; RRID: AB_11178526.

Techniques: Immunostaining, Recombinant, Luciferase, Reporter Assay, Plasmid Preparation, Purification, Fractionation, cDNA Synthesis, Mutagenesis, Isolation, Transfection, Generated, Software