Journal: Oxidative medicine and cellular longevity

Article Title: Mitochondrial-Targeting Antioxidant SS-31 Suppresses Airway Inflammation and Oxidative Stress Induced by Cigarette Smoke.

doi: 10.1155/2021/6644238

Figure Lengend Snippet: Figure 6: SS-31 prevents mitochondrial dysfunction in CS-induced mouse lung. (a–d) Protein levels of OPA1, MFF (a, b), and cytochrome c (c, d) in lungs were evaluated by western blot following exposure to CS with or without SS-31 pretreatment. GAPDH or COX IV was used as a loading control (n = 3 per group). ∗∗P < 0:01 and ∗∗∗P < 0:001 vs. control; #P < 0:05 vs CS. Abbreviations: COX IV: cytochrome c oxidase IV; CS: cigarette smoke; Cyt c: cytochrome c; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; MFF: mitochondrial fission factor; mit: mitochondrion; OPA1: optic atrophy 1; SS-31 (H): high-dose SS-31 (5 mg/kg); SS-31 (L): low-dose SS-31 (2.5 mg/kg).

Article Snippet: After blocking with 5% bovine serum albumin in Trisbuffered saline at room temperature for 1 h, the membrane was incubated overnight at 4°C with antibodies against optic atrophy (OPA) 1 (mouse antibody from Cell Signaling Technology; human antibody from Proteintech, Rosemont, IL, USA); mitochondrial fission factor (MFF) (mouse antibody from Cell Signaling Technology; human antibody from Proteintech); and extracellular signal-regulated kinase (ERK), phosphorylated- (P-) ERK, P38, P-P38, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (all from Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: Oxidative medicine and cellular longevity

Article Title: Mitochondrial-Targeting Antioxidant SS-31 Suppresses Airway Inflammation and Oxidative Stress Induced by Cigarette Smoke.

doi: 10.1155/2021/6644238

Figure Lengend Snippet: Figure 11: SS-31 protects against CSE-induced mitochondrial dysfunction via suppression of MAPK signaling. (a, b) Protein levels of OPA1 and MFF detected by western blot in BEAS-2B cells (n = 3 per group). ∗∗P < 0:01, ∗∗∗P < 0:001, and ∗∗∗∗P < 0:0001 vs. control; #P < 0:05 and ##P < 0:01 vs. ani+CSE; &P < 0:05 vs. CSE. Abbreviations: ani: anisomycin; CSE: cigarette smoke extract; MFF: mitochondrial fission factor; OPA1: optic atrophy 1.

Article Snippet: After blocking with 5% bovine serum albumin in Trisbuffered saline at room temperature for 1 h, the membrane was incubated overnight at 4°C with antibodies against optic atrophy (OPA) 1 (mouse antibody from Cell Signaling Technology; human antibody from Proteintech, Rosemont, IL, USA); mitochondrial fission factor (MFF) (mouse antibody from Cell Signaling Technology; human antibody from Proteintech); and extracellular signal-regulated kinase (ERK), phosphorylated- (P-) ERK, P38, P-P38, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (all from Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nitric oxide modulates macrophage responses to M. tuberculosis infection through activation of HIF-1α and repression of NF-kB

doi: 10.4049/jimmunol.1700515

Figure Lengend Snippet: (A) Western blots 24h post-infection from whole cell lysates and nuclear extracts for RelA, RelB, NF-kB1, and IkBa, with tubulin as a cytoplasmic marker and Histone H3 as a nuclear marker. Western blots with wildtype and Nos2−/− BMDM either uninfected [un] or M. tuberculosis infected with IFN-γ activation [TB/G]. For nuclear extracts, [TB/G] performed in biological duplicate for each repeat. (B) Confocal microscopy at 63x for RelA 24h post-infection in wildtype and Nos2−/− BMDM, uninfected or during M. tuberculosis infection with IFN-γ activation [TB/G]. For microscopy, the M. tuberculosis strain Erdman, carrying a fluorescent mCherry was utilized. (C) Quantification of p65 positive nuclei from the same samples as in (B), but from 20x images. >800 nuclei were analyzed for each condition, error bars represent SD between 5 fields. (D) RNAseq data showing number of reads for the IL1R antagonist (Ilrn) and IL1R (Il1r1) in wildtype BMDM infected with M. tuberculosis [TB] or M. tuberculosis with IFN-γ activation [TB/G]. (E) qPCR for Il1b transcript in wildtype and Il1r−/− BMDM, either untreated [un], during M. tuberculosis infection with IFN-γ activation [TB/G], or during M. tuberculosis infection with IFN-γ activation and 1400W treatment at 25uM [TB/G+1400W]. All data representative of 2 or more experiments. The p values were determined using an unpaired t test. **p<.01, ***p<.001

Article Snippet: The following primary antibodies were used: HIF-1α (NB100-479, Novus Biologicals), IL-1b (AF-401-NA, R&D systems), and the following Cell Signaling Technology antibodies: HIF-1α (D2U3T), RelA (D14E12), RelB (C1E4), NF-kB1 (D4P4D), IkBa (L35A5), α/β-Tubulin (2148), Histone H3 (D1H2), and β-actin (13E5).

Techniques: Western Blot, Infection, Marker, Activation Assay, Confocal Microscopy, Microscopy