Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial NDUFA4L2 attenuates the apoptosis of nucleus pulposus cells induced by oxidative stress via the inhibition of mitophagy

doi: 10.1038/s12276-019-0331-2

Figure Lengend Snippet: After pretreatment with FCCP or CsA, primary nucleus pulposus cells were exposed to 400 μM TBHP for 6 h. a Western blotting for the protein levels of cleaved caspase-3, Bcl-2/Bax, Beclin-1, Parkin and LC-3II. b–f Quantitative analysis of the protein content of cleaved caspase-3, Bcl-2/Bax, Beclin-1, Parkin and LC-3II. g Flow cytometry was used to detect apoptosis in nucleus pulposus cells. h Quantitative analysis of flow cytometry for the detection of apoptosis. i Fluorescence images of NP cells infected with mRFP-GFP-LC-3 adenovirus. sensGFP is sensitive to the pH changes owing to the fusion of autophagosomes and lysosomes, whereas mRFP is stable. When autophagy was induced, autophagosomes and lysosomes were fused, sensGFP was quenched and mRFP was increased. j Hoechst 33258 staining detected apoptosis in the cell nucleus. Nuclear condensation was observed in apoptotic cells. The data are presented as the mean ± S.D. *** p < 0.001, ** p < 0.01, * p < 0.05 ( n = 5). Cd caspase-3: cleaved caspase-3.

Article Snippet: The membranes were blocked with TBST-buffered saline solution containing 5% dry milk for 2 h and then incubated overnight at 4 °C or at 25 °C for 2 h with primary antibodies against HIF-1α (1:1000, Proteintech, China), Bcl-2 (1:1000, Proteintech, China), Bax (1:1000, Proteintech, China), Beclin-1 (1:1000, CST, USA), LC-3 (1:1000, Novus, USA), cleaved caspase-3 (1:1000, Proteintech, China), cleaved PARP (1:1000, CST, USA), NDUFA4L2 (1:1000, Proteintech, China), and P62 (1:1000, Proteintech, China).

Techniques: Western Blot, Flow Cytometry, Fluorescence, Infection, Staining

Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial NDUFA4L2 attenuates the apoptosis of nucleus pulposus cells induced by oxidative stress via the inhibition of mitophagy

doi: 10.1038/s12276-019-0331-2

Figure Lengend Snippet: Primary nucleus pulposus cells were transfected with si-NDUFA4L2, si-NC, pc-NDUFA4L2 or pc-NC for 48 h and then exposed to TBHP. a Western blotting for the protein levels of NDUFA4L2, LC-3II and Parkin was performed to determine the function of si-NDUFA4L2. b Western blotting for the protein levels of NDUFA4L2, LC-3II and Parkin was performed to determine the function of pc-NDUFA4L2. c Western blotting for the protein levels of NDUFA4L2, LC-3II, Parkin and Bcl-2/Bax was performed to investigate the function of NDUFA4L2 in NP cells exposed to TBHP. d–f Quantitative analysis of the protein content of P62, Bcl-2/Bax and LC-3II. g Immunofluorescence images showing staining for NDUFA4L2 (red), cytochrome C (green), and DAPI (blue) and merged images of two signals. h Immunofluorescence images showing staining for NDUFA4L2 (red), LC-3 (green), and DAPI (blue) and merged images of two signals. The data are presented as the mean ± S.D. *** p < 0.001, ** p < 0.01, * p < 0.05 ( n = 5).

Article Snippet: The membranes were blocked with TBST-buffered saline solution containing 5% dry milk for 2 h and then incubated overnight at 4 °C or at 25 °C for 2 h with primary antibodies against HIF-1α (1:1000, Proteintech, China), Bcl-2 (1:1000, Proteintech, China), Bax (1:1000, Proteintech, China), Beclin-1 (1:1000, CST, USA), LC-3 (1:1000, Novus, USA), cleaved caspase-3 (1:1000, Proteintech, China), cleaved PARP (1:1000, CST, USA), NDUFA4L2 (1:1000, Proteintech, China), and P62 (1:1000, Proteintech, China).

Techniques: Transfection, Western Blot, Immunofluorescence, Staining

Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial NDUFA4L2 attenuates the apoptosis of nucleus pulposus cells induced by oxidative stress via the inhibition of mitophagy

doi: 10.1038/s12276-019-0331-2

Figure Lengend Snippet: Primary nucleus pulposus cells were transfected with si-NDUFA4L2 or si-NC for 48 h and then exposed to FCCP or CsA. a Western blotting for the protein levels of NDUFA4L2, cleaved caspase-3, P62, Parkin and LC-3II. b – f Quantitative analysis of the protein content of NDUFA4L2, cleaved caspase-3, P62, Parkin and LC-3II. g Fluorescence images of NP cells infected with mRFP-GFP-LC-3 adenovirus. sensGFP is sensitive to the pH changes due to the fusion of autophagosomes and lysosomes, whereas mRFP is stable. When autophagy was induced, autophagosomes and lysosomes were fused, sensGFP was quenched and mRFP was increased. The data are presented as the mean ± S.D. *** p < 0.001, ** p < 0.01, * p < 0.05 ( n = 5).

Article Snippet: The membranes were blocked with TBST-buffered saline solution containing 5% dry milk for 2 h and then incubated overnight at 4 °C or at 25 °C for 2 h with primary antibodies against HIF-1α (1:1000, Proteintech, China), Bcl-2 (1:1000, Proteintech, China), Bax (1:1000, Proteintech, China), Beclin-1 (1:1000, CST, USA), LC-3 (1:1000, Novus, USA), cleaved caspase-3 (1:1000, Proteintech, China), cleaved PARP (1:1000, CST, USA), NDUFA4L2 (1:1000, Proteintech, China), and P62 (1:1000, Proteintech, China).

Techniques: Transfection, Western Blot, Fluorescence, Infection

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Poloxamer 407 and Hyaluronic Acid Thermosensitive Hydrogel-Encapsulated Ginsenoside Rg3 to Promote Skin Wound Healing

doi: 10.3389/fbioe.2022.831007

Figure Lengend Snippet: Effect of Rg3-Gel on the expression of various proteins in wound tissues. (A) Representative images of pan-keratin immunofluorescence staining on day 16 post surgery. (B) Expression of pan-keratin protein affected through the Rg3-Gel treatment in skin tissue in mouse (# p < 0.05, ## p < 0.01, and n = 3). (C) Representative western blotting of Akt, JNK, ERK, p38, and their phosphorylated proteins in skin tissue of three groups. (D) Effects of different treatment on the protein expressions of NF-κB, (E) p62, (F) Beclin-1, and LC3 (# p < 0.05, ## p < 0.01, and n = 3). Analysis of the MAPK/NF-κB signaling pathway and expression of autophagy-related proteins.

Article Snippet: Ginsenoside Rg3 was produced in the laboratory (the purified Rg3, having two configurations >95% total composition); P407 was supplied by BASF (Ludwigshafen, Germany); low-molecular-weight chitosan obtained from shrimp shells ( Pandalus borealis ) was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd (the deacetylation degree of chitosan was 95.8%); sodium hyaluronate (15–25 MDa) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd; SDS was purchased from Beijing Solarbio Science Technology Co., Ltd.; HPLC-grade acetonitrile (LiChrosolv ® , CAS-No: 67-56-1) was purchased from Merck (Darmstadt, Germany); RIPA lysis buffer and BCA protein assay kit (BCA) were purchased from Beyotime Institute of Biotechnology (Jiangsu, China); the antibodies against p-ERK, p-JNK, and p-p38 were obtained from Cell Signaling Technology (Beverly, United States); antibodies against β-actin, GAPDH, and the goat antirabbit secondary antibody, NF-κB p65, Akt, ERK, JNK, p38, LC3, and Beclin-1 were obtained from the Proteintech (Santa Cruz, CA, United States); and antibodies against p-Akt and p62 were obtained from Arigo Biolaboratories Corp.

Techniques: Expressing, Immunofluorescence, Staining, Western Blot

Journal: American Journal of Physiology - Renal Physiology

Article Title: The mechanosensitive BKα/β1 channel localizes to cilia of principal cells in rabbit cortical collecting duct (CCD)

doi: 10.1152/ajprenal.00256.2016

Figure Lengend Snippet: Antibodies (Abs) used for immunoperfusion

Article Snippet: Goat IgG anti-aquaporin-2 (C-17) , 1:100 , Santa Cruz Biotechnology (sc-9882) , A488-rabbit IgG anti-goat IgG , 1:500 , Molecular Probes ( {"type":"entrez-nucleotide","attrs":{"text":"A11078","term_id":"490929","term_text":"A11078"}} A11078 ).

Techniques: