Journal: Oxidative medicine and cellular longevity

Article Title: Inhibition of α -Synuclein Accumulation Improves Neuronal Apoptosis and Delayed Postoperative Cognitive Recovery in Aged Mice.

doi: 10.1155/2021/5572899

Figure Lengend Snippet: Figure 3: Peripheral surgery increased mitochondrial α-syn accumulation and damage. (a) The hippocampus was stained to determine colocalization of α-syn (red) and COXIV (an inner mitochondrial membrane (green)). Cell nuclei were stained with DAPI (blue). Arrows point to mitochondrial α-syn; scale bars = 10 μm, n = 5. (b) Transmission electron microscopy showing mitochondrial morphology in the hippocampus after surgery (n = 3 mice per group) and assessment of abnormal cristae (n = 57 −60 cells). Scale bars = 500 nm. Results are presented as mean ± SEM (one-way analysis of variance). ∗∗∗P < 0:001, control (Con) or anesthesia-alone (Ane) vs. surgery plus anesthesia (Sur).

Article Snippet: Accumulation of SNCA oligomers was detected by immunohistochemistry using a rabbit polyclonal SNCA oligomer-specific Syn33 antibody (ABN2265; Merck Millipore, Billerica, MA, USA), a mouse monoclonal antibody against total SNCA (4D6, ab1903; Abcam), a rabbit polyclonal COXIV antibody (11242-1-AP; Proteintech), a rabbit monoclonal antibody against p-α-syn (Ser129, ab1903; Abcam), and a rabbit monoclonal antibody against Iba-1 (Wako 019-19741; Rosemont, IL, USA).

Techniques: Staining, Membrane, Transmission Assay, Electron Microscopy, Control

Journal: Cell Death & Disease

Article Title: Elevated SFXN2 limits mitochondrial autophagy and increases iron-mediated energy production to promote multiple myeloma cell proliferation

doi: 10.1038/s41419-022-05272-z

Figure Lengend Snippet: a WB examined the expressions of autophagy/mitophagy-related proteins ATG5, ATG7, LC3, PINK1, and Parkin in Ctrl and SFXN2-KD cells. b Representative transmission electron microscopic images of mitochondrial morphology indicated by red arrows in Ctrl and SFXN2-KD cells. c Quantification of numbers and sizes of mitochondria by ImageJ in Ctrl and SFXN2-KD cells ( p < 0.05, using Kruskal Wallis test in R). d IF staining of TRITC-labeled LC3b in WT and SFXN2-OE ARP1 and H929 cells. Quantification of fluorescence intensity of TRITC-labeled LC3b in WT and SFXN2-OE ARP1 and H929 cells ( n = 3 in every group, * p < 0.05, ** p < 0.01). e Representative photographs showed autophagic vesicles indicated by red arrows in WT and SFXN2-OE ARP1 and H929 cells post EBSS treatment. Scale bars: 0.5 μm. Quantification of starvation-induced autophagic vesicles in WT and SFXN2-OE cells ( n = 3 in each group, * p < 0.05). f WB examined the autophagy-related proteins ATG5, ATG7, Beclin1, LC3, PINK1, and Parkin in EBSS-treated WT and SFXN2-OE ARP1 cells compared to non-treated cells. g Transmission electron microscope testified representative mitochondrial morphology indicated by red arrows in WT and SFXN2-OE cells. h Numbers and sizes of mitochondria were analyzed by ImageJ ( p < 0.05, using Kruskal Wallis test in R). i Relative mtDNA content of WT, SFXN2-OE, SFXN2-Ctrl, and SFXN2-KD cells were measured by a competitive PCR method. j The level of ATP in WT, SFXN2-OE, SFXN2-Ctrl, and SFXN2-KD cells were determined by the ATP Bioluminescence Assay Kit (* p < 0.05, *** p < 0.001).

Article Snippet: The following commercial antibodies were used in this study: SFXN2 (ab67191), TOMM20 (ab186734) from Abcam; HO1 (66743-1-Ig), PINK-1(23274-1-AP), PARKIN (14060-1-AP), and P62/SQSTM1 (18420-1-AP) from ProteinTech; Atg7 (#2631s), Beclin-1 (#3738), LC3A/B (#12741), α-tubulin (#2125s), β-actin (#3700s), and goat anti-rabbit IgG-HRP (#7074) from Cell Signaling Technology; Atg5 (Biological, 110-53818), COX4 (Bioss, bsm-33037M), goat anti-mouse IgG-HRP (SANTA, SC-2005).

Techniques: Transmission Assay, Staining, Labeling, Fluorescence, Microscopy, ATP Bioluminescent Assay

Journal: Cell Death & Disease

Article Title: Elevated SFXN2 limits mitochondrial autophagy and increases iron-mediated energy production to promote multiple myeloma cell proliferation

doi: 10.1038/s41419-022-05272-z

Figure Lengend Snippet: a Co-IP/MS identified two original peptide sequences corresponding to HO1 (unipro ID: P09601). b IP assay showed the interaction between SFXN2 and HO1. c Luciferase activity driven by a LC3B promoter sequence screened potent molecules influencing autophagic activity in WT and SFXN2-OE ARP1 cells before and after treatment of HO1 inhibitor (HO-1-IN-1). d WB analysis showed the expressions of mitophagy-related proteins ATG7, ATG5, Parkin, PINK1, and LC3 in ARP1 cells treated with EBSS and HO-1-IN-1 individually or both. e ARP1 cells transduced with mCherry-eGFP-LC3 were incubated with either complete media or EBSS for 48 h. All the images were captured with the confocal microscope (mCherry-LC3, Red; eGFP-LC3, Green). f , g WB tested the expressions of HO1 and SFXN2 in SFXN2-OE ( f ) and SFXN2-KD ( g ) cells. h Flow cytometry analysis examined intracellular ROS levels in WT and SFXN2-OE cells treated with HO-1-IN-1(left panel). Quantification analysis for fluorescence intensity was performed in WT and SFXN2-OE cells (right panel).

Article Snippet: The following commercial antibodies were used in this study: SFXN2 (ab67191), TOMM20 (ab186734) from Abcam; HO1 (66743-1-Ig), PINK-1(23274-1-AP), PARKIN (14060-1-AP), and P62/SQSTM1 (18420-1-AP) from ProteinTech; Atg7 (#2631s), Beclin-1 (#3738), LC3A/B (#12741), α-tubulin (#2125s), β-actin (#3700s), and goat anti-rabbit IgG-HRP (#7074) from Cell Signaling Technology; Atg5 (Biological, 110-53818), COX4 (Bioss, bsm-33037M), goat anti-mouse IgG-HRP (SANTA, SC-2005).

Techniques: Co-Immunoprecipitation Assay, Luciferase, Activity Assay, Sequencing, Transduction, Incubation, Microscopy, Flow Cytometry, Fluorescence