mouse anti heat shock protein 60 hsp60  (Santa Cruz Biotechnology)

Journal: Experimental Physiology

Article Title: Effect of prolonged voluntary wheel running on oxidative stress and defence mechanisms in cortex and hippocampus of healthy female rats

doi: 10.1113/ep092815

Figure Lengend Snippet: FIGURE 6 Mitochondrial BER. (a) Representative immunoblots of target proteins in mitochondrial extracts from cortex and hippocampus. Proteins probed for are shown on the left. APE1, apurinic/apyrimidinic endonuclease 1; HSP60, heat shock protein 60. (b) Representative gel from in vitro AP endonuclease activity assay with mitochondrial extracts from hippocampus. S, substrate; P, product; AP, oligonucleotide with AP site; C, control oligonucleotide without a lesion. (c–f) All values are normalized to HSP60 (mitochondrial loading control) and set relative to the sedentary group mean. Bars represent means ± SD. Differences were evaluated by Student’s unpaired t-test (c) or Mann–Whitney U-test (d–f). (c) n = 10 and 8 for sedentary and running group, respectively. (d) n = 10 and 7 for sedentary and running group, respectively. (e) n = 9 and 8 for sedentary and running group, respectively. (f) n = 7 and 9 for sedentary and running group, respectively. One outlier was excluded from the analysis in the sedentary group for (e, f). Other discrepancies in sample size are due to technical issues in assays or insufficient yield from mitochondrial purification. BER, base excision repair.

Article Snippet: anti-8-oxoguanine DNA glycosylase 1 (OGG1) (Novus Biologicals, cat. no. NB100-106, 1:500, RRID: AB_10104097), mouse anti-superoxide dismutase 1 (SOD1) (Santa Cruz Biotechnology, Dallas, TX, USA, cat. no. sc-101523, 1:500, RRID: AB_2191632), mouse anti-superoxide dismutase 2 (SOD2) (Santa Cruz Biotechnology, cat. no. sc-137254, 1:200, RRID: AB_2191808), mouse anti-catalase (CAT) (Santa Cruz Biotechnology, cat. no. sc-271803, 1:500, RRID: AB_10708550), mouse anti-heat shock protein 60 (HSP60) (Santa Cruz Biotechnology, cat. no. sc-271215, 1:1000, RRID: AB_10607973), and mouse antiβ-actin (Sigma-Aldrich, St Louis, MO, USA, cat. no. A2228, 1:10,000, RRID: 476697).

Techniques: Western Blot, In Vitro, Activity Assay, Control, MANN-WHITNEY, Purification

Journal: Frontiers in Endocrinology

Article Title: 17β-Estradiol Induces Mitophagy Upregulation to Protect Chondrocytes via the SIRT1-Mediated AMPK/mTOR Signaling Pathway

doi: 10.3389/fendo.2020.615250

Figure Lengend Snippet: 17β-E2 treatment upregulated the phosphorylation level of LC3, TOM20, and Hsp60 in ATDC5 chondrocytes. The chondrocytes were pretreated with 15 μM NAM or 20 μM Compound C for 30 min and then incubated with or without 1 × 10 -7 M 17β-E2 for 24 h. The phosphorylation levels of (A) microtubule-associated protein 1A/1B light chain 3 (LC3), (B) translocase of outer membrane 20 (TOM20), (C) heat shock protein 60 (Hsp60), and β-actin protein were detected by Western blot analysis. These experiments were independently repeated 3 times. * P < 0.05 versus the control group indicated a significant difference.

Article Snippet: The polyclonal goat anti-mouse heat shock protein 60 (Hsp60) antibody (sc-1052) (1:200), the monoclonal mouse anti-mouse microtubule-associated protein 1A/1B-light chain 3 (LC3) antibody (sc-376404) (1: 100), rabbit monoclonal anti-β-actin and the polyclonal rabbit anti-mouse primary translocase of outer membrane (TOM) 20 antibody (sc-11415) (1: 200) were purchased from Sigma (MO, USA).

Techniques: Incubation, Western Blot

Journal: mSphere

Article Title: Development of a Plasmid Shuttle Vector System for Genetic Manipulation of Chlamydia psittaci

doi: 10.1128/mSphere.00787-20

Figure Lengend Snippet: GFP expression and mCherry induction in pCps-Tet-mCherry-transformed C. psittaci strain 02DC15. Transformed C. psittaci 02DC15 bacteria were grown in epithelial cells with 1U/ml PEN for 24 and 48 h. (A) GFP fluorescence of chlamydial inclusions was visualized in living cells without fixing and staining. aTC (10 and 100 ng/ml) was added at 1 hpi to induce mCherry expression. Images are representative of three independent experiments. White arrows show chlamydial inclusions. White scale bars represent 10 μm. (B) mCherry was analyzed by Western blotting and densitometric analyses at 24 and 48 hpi. mCherry protein amounts were normalized to chlamydial HSP60. GAPDH was used as a loading control. ( n = 4; mean ± SEM; Sidak’s multiple comparison: ***, P ≤ 0.001).

Article Snippet: Rabbit anti-mCherry and mouse anti-heat shock protein 60 (HSP60) were purchased from Thermo Fisher Scientific (Waltham, MA).

Techniques: Expressing, Transformation Assay, Fluorescence, Staining, Western Blot