Journal: Experimental and therapeutic medicine

Article Title: Xanthohumol attenuates renal ischemia/reperfusion injury by inhibiting ferroptosis.

doi: 10.3892/etm.2023.12269

Figure Lengend Snippet: Figure 2. XN prevents ferroptosis during RIRI. (A) ROS production in kidney tissue was measured by the dihydroethidium fluorescence probe; magnifica‑ tion, x200. (B) Quantitative analysis of ROS levels. (C) SOD, (D) MDA and (E) GSH levels in kidney tissues of rats. (F) Western blotting of (G) GPX4 and (H) ACSL4 in kidney tissues. *P<0.05 vs. sham, #P<0.05 vs. IR and &P<0.05 vs. XN + IR. XN, xanthohumol; RIRI, renal ischemia/reperfusion injury; ROS, reactive oxygen species; SOD, superoxide dismutase; MDA, malondialdehyde; GSH, glutathione; ACSL4, long‑chain acyl‑CoA synthetase.

Article Snippet: Membranes were incubated with primary anti‐ bodies against GPX4 (1:1,000; cat. no. 67763‐1‐Ig; Proteintech Group, inc.), long‐chain acyl‐CoA synthetase (ACSl4; 1:1,000; cat. no. 22401‐1‐AP; Proteintech Group, Inc.), Nrf2 (1:1,000; cat. no. 80593‐1‐RR; Proteintech Group, Inc.), heme oxygenase (HO)‐1 (1:1,000; cat. no. 10701‐1‐AP; Proteintech Group, Inc.) or GAPDH (1:1,000; cat. no. 60004‐1‐Ig; Proteintech Group, inc.) overnight at 4 ̊C following blocking with 5% BSA for 2 h at room temperature (Beijing Solarbio Science & Technology Co., ltd.).

Techniques: Fluorescence, Western Blot

Journal: Experimental and therapeutic medicine

Article Title: Xanthohumol attenuates renal ischemia/reperfusion injury by inhibiting ferroptosis.

doi: 10.3892/etm.2023.12269

Figure Lengend Snippet: Figure 3. XN inhibits HR‑induced ferroptosis in HK‑2 cells. (A) Cell Counting Kit‑8 was used to evaluate cell viability. (B) SOD, (C) MDA and (D) GSH levels in HK‑2 cells. (E) Western blotting of (F) GPX4 and (G) ACSL4 in HK‑2 cells. *P<0.05 vs. control, #P<0.05 vs. HR and &P<0.05 vs. XN + HR. XN, xantho‑ humol; HR, hypoxia‑reoxygenation; SOD, superoxide dismutase; MDA, malondialdehyde; GSH, glutathione; prot, protein; GPX4, glutathione peroxidase 4; ACSL4, long‑chain acyl‑CoA synthetase.

Article Snippet: Membranes were incubated with primary anti‐ bodies against GPX4 (1:1,000; cat. no. 67763‐1‐Ig; Proteintech Group, inc.), long‐chain acyl‐CoA synthetase (ACSl4; 1:1,000; cat. no. 22401‐1‐AP; Proteintech Group, Inc.), Nrf2 (1:1,000; cat. no. 80593‐1‐RR; Proteintech Group, Inc.), heme oxygenase (HO)‐1 (1:1,000; cat. no. 10701‐1‐AP; Proteintech Group, Inc.) or GAPDH (1:1,000; cat. no. 60004‐1‐Ig; Proteintech Group, inc.) overnight at 4 ̊C following blocking with 5% BSA for 2 h at room temperature (Beijing Solarbio Science & Technology Co., ltd.).

Techniques: CCK-8 Assay, Western Blot, Control

Journal: Experimental and therapeutic medicine

Article Title: Xanthohumol attenuates renal ischemia/reperfusion injury by inhibiting ferroptosis.

doi: 10.3892/etm.2023.12269

Figure Lengend Snippet: Figure 5. XN inhibits HR‑induced ferroptosis via the Nrf2/HO‑1 pathway. (A) HK‑2 cells were transfected with si‑Nrf2 or si‑con before HR and XN; western blotting was used to examine Nrf2 in cells. (B) Quantitative analysis of Nrf2 expression. (C) Cell Counting Kit‑8 was used to evaluate cell viability. (D) SOD, (E) MDA and (F) GSH levels in HK‑2 cells. (G) Western blotting of (H) Nrf2, (I) HO‑1, (J) GPX4 and (K) ACSL4 in HK‑2 cells. (L and M) ROS production in HK‑2 cells was measured by the 2,7‑dichlorofluorescein diacetate fluorescent probe; magnification, x400. *P<0.05 vs. si‑con, #P<0.05 vs. si‑con + HR, &P<0.05 vs. XN + si‑con + HR. si‑con, negative control small interfering RNA; XN, xanthohumol; ROS, reactive oxygen species; HR, hypoxia‑reoxygenation; SOD, superoxide dismutase; prot, protein; MDA, malondialdehyde; GSH, glutathione; GPX4, glutathione peroxidase 4; ACSL4, long‑chain acyl‑CoA synthetase.

Article Snippet: Membranes were incubated with primary anti‐ bodies against GPX4 (1:1,000; cat. no. 67763‐1‐Ig; Proteintech Group, inc.), long‐chain acyl‐CoA synthetase (ACSl4; 1:1,000; cat. no. 22401‐1‐AP; Proteintech Group, Inc.), Nrf2 (1:1,000; cat. no. 80593‐1‐RR; Proteintech Group, Inc.), heme oxygenase (HO)‐1 (1:1,000; cat. no. 10701‐1‐AP; Proteintech Group, Inc.) or GAPDH (1:1,000; cat. no. 60004‐1‐Ig; Proteintech Group, inc.) overnight at 4 ̊C following blocking with 5% BSA for 2 h at room temperature (Beijing Solarbio Science & Technology Co., ltd.).

Techniques: Transfection, Western Blot, Expressing, CCK-8 Assay, Negative Control, Small Interfering RNA

Journal: Molecules

Article Title: Label-Free Quantitative Proteomics to Explore the Action Mechanism of the Pharmaceutical-Grade Triticum vulgare Extract in Speeding Up Keratinocyte Healing

doi: 10.3390/molecules27031108

Figure Lengend Snippet: List of the significantly quantified proteins whose “cellular component” is annotated to be “extracellular”. Each protein is reported with its accession, gene name, TVE vs. control abundance ratio, and the related p -value.

Article Snippet: The membrane was soaked for 1 h at room temperature in 5% non-fat dried milk containing TBS-t solution (31 mM Tris, pH 8, 170 mM NaCl, 3.35 mM KCl, 0.05% Tween 20) and then incubated overnight at 4 °C with a monoclonal primary anti-body against fibulin 1 (FBLN1, mouse, 1:500 v/v , sc-25281, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), tissue plasminogen A (tPA, 1:250 v/v , sc-69740, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), kallikrein 6 (KLK6, 1:250 v/v , sc-374564, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), and integrin β1 (ITGβ1, 1:1000 v/v , 34971, Cell Signaling Technologies, Danvers, Massachusetts), all in milk/TBS-t. After recognition of the primary antibodies from the proper HPR-tagged secondary ones (1:500 v/v mouse for FBLN1, tPA, and KLK6 and 1:1500 v/v rabbit for ITGβ1), chemiluminescence, developed upon treatment of the membrane with a mixture of luminol and hydrogen peroxide, was detected using an ImageQuant LAS4000 imaging system (GE Healthcare, Waukesha, WI, USA).

Techniques: Control, Coagulation

Journal: PLoS ONE

Article Title: p38 MAPK Signaling in Postnatal Tendon Growth and Remodeling

doi: 10.1371/journal.pone.0120044

Figure Lengend Snippet: Immunoblot and immunohistochemistry detection reagents used in this study.

Article Snippet: Slides were incubated with primary anti- bodies against rabbit anti-Ki67 (Abcam), rabbit anti-CD146 (Abcam), biotinylated rabbit anti-F4/80 (Abcam), rabbit anti-procollagen type I (Santa Cruz Biotechnology, Dallas, TX), or biotinylated hyaluronic acid binding protein (HABP, Millipore, Billerica, MA).

Techniques: Western Blot, Immunohistochemistry, Binding Assay

Journal: Biochemical Journal

Article Title: Regulation of mitochondrial biogenesis in brown adipose tissue: nuclear respiratory factor-2/GA-binding protein is responsible for the transcriptional regulation of the gene for the mitochondrial ATP synthase β subunit

doi: 10.1042/bj3310121

Figure Lengend Snippet: Figure 6 Expression of chimaeric constructs containing the ®306/®266 region upstream of the basal TK promoter in non-differentiated and differentiated HIB-1B brown adipocytes: effects of two double-point mutations suppressing the binding to NRF2/GABP or Sp1

Article Snippet: When antibodies were used, 1 μl of anti-sera against GA-binding protein (GABP) β1 or Sp3, gifts from Dr. S. McKnight (Tularik Inc., San Francisco, U.S.A.) and Dr. G. Suske (University of Marburg, Germany) respectively, or 1 μl of anti-bodies against Sp1 or retinoic acid receptor α (Santa Cruz Inc.) were included in the incubation mixture.

Techniques: Expressing, Construct, Binding Assay