Journal:

Article Title: The function of Xenopus Bloom's syndrome protein homolog (xBLM) in DNA replication

doi:

Figure Lengend Snippet: Characterization of xBLM protein. (A) Coomassie blue staining of the purified His-tagged recombinant xBLM protein. (B) Helicase activity of the recombinant xBLM. The substrate is a 5′-32P-labeled 48mer oligonucleotide hybridized to m13mp18 single-stranded DNA. The purified xBLM protein was incubated with the substrate in the presence of either buffer alone or nucleotides (ATP or dATP). The right lane contains the heat-denatured substrate. The dissociated oligonucleotide was separated from m13 on 12% polyacrylamide gels. (C) Western blot of interphase egg extracts (2 μL) probed with the affinity-purified rabbit anti-xBLM antibody and detected by ECL (Amersham). The 160-kD band is xBLM. The small band is not always detected, suggesting that it is a degradation product. (D) Immunodepletion of xBLM from egg extracts. Supernatants (2 μL) from the extracts that had been depleted with either anti-xBLM or control antibodies were analyzed by Western blot with the affinity-purified rabbit anti-xBLM antibody. Molecular markers are labeled as kilodaltons on the right side of panels A, C, and D.

Article Snippet: Immunodepletion was performed by incubating extracts (diluted twofold with egg lysis buffer) with Affi-gel Protein A beads (Bio-Rad) that had been precoated with either the affinity-purified rabbit anti-xBLM antibody or rabbit IgG.

Techniques: Staining, Purification, Recombinant, Activity Assay, Labeling, Incubation, Western Blot, Affinity Purification

Journal: Development, Growth & Differentiation

Article Title: BMP ‐Dependent Modulation of ROS Generation and Scavenging Controls Interdigital Cell Death

doi: 10.1111/dgd.70034

Figure Lengend Snippet: BMP signaling promotes ROS production in the interdigital regions. (a) Cell death (LysoTracker, green) and ROS (DHE, red) were detected in the hindlimbs of chicken embryos 10–17 h after implantation of heparin‐acrylic beads (asterisks) soaked with PBS ( n = 5) or BMP2 ( n = 7) at HH28‐29. Note that cell death and ROS signals increased around BMP2‐soaked beads (arrowheads). (b) Quantification of LysoTracker‐ or DHE‐positive puncta in ID3 of the left and right hindlimbs indicated in (a). Each pair of symbols represents an individual embryo. Statistical analysis was performed using the Wilcoxon matched‐pairs signed‐rank test, and q values are reported. (c) Cell death (LysoTracker, green) and ROS (DHE, red) signals were detected 13–17 h after implantation of Affi‐Gel Blue Gel beads (asterisks) soaked with PBS ( n = 8) or Noggin ( n = 9) at HH29‐30. Note that cell death and ROS signals were diminished around Noggin‐soaked beads (bracket). (d) Quantification of LysoTracker‐ or DHE‐positive puncta in ID3 of the left and right limbs shown in (c). Each pair of symbols represents an individual embryo. Statistical analysis was performed using the Wilcoxon matched‐pairs signed‐rank test, and q values are reported. Scale bars, 500 μm. ID3, interdigital region 3.

Article Snippet: CM Affi‐Gel Blue Gel beads (153–7304, BioRad, California) were washed, dried, and soaked in Noggin or PBS.

Techniques: