α sod2  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: A mitochondrial quality control mechanism reverses the phagosome maturation arrest caused by Mycobacterium tuberculosis

doi: 10.1101/2023.12.01.569475

Figure Lengend Snippet: (A) High-resolution images of immunostained TOM20 (green) in H37Rv-infected WT and p62 KD macrophages at 24 hpi. The zoomed images on the right show encircled TOM20 + -MDVs in the p62 KD cells. (B) High-resolution images of immunostained TOM20 (red) and MCU (green). The encircled area depicts TOM20 + MDVs in the zoomed images at the right. (C) Quantification of number of TOM20 + MDVs in uninfected and Mtb -infected WT and p62 KD cells. Data show mean ± SD, n=6 fields having 5-10 cells per field, from two independent experiments (D) Schematic depiction of the presence of different proteins on mitochondrial matrix, inner and outer membrane. (E) Bar graphs show quantitative interactions of matrix mitochondrial protein spots of PDHA1, HSP60 and SOD2 with TOM20 structures. (F) Bar graphs show the quantitative association of inner membrane mitochondrial protein spots of ECSIT and MCU with TOM20 structures. ( G - K ) Immunofluorescent images depict the interaction of mCherryH37Rv with ECSIT (G), MCU (H ), PDHA1 (I) , SOD2 (J) and HSP60 (K) (green), respectively, in WT and p62 KD THP-1 macrophages at 24 hpi. The 3D reconstruction shows H37Rv structures and indicates mitochondrial protein spots. The yellow spots are <0.2 μm from H37Rv structures, and the pink spots are >0.2 μm from bacterial structures. The bar graph on the right shows a number of mitochondrial protein spots on Mtb . Data show (E-K) mean ± SD, n≥5 fields having 20-35 cells per field, from two independent experiments, Scale bar =10 μm unless specified.

Article Snippet: The primary antibodies used were: α-p62/SQSTM1 (Abcam, ab155686), α-OPTN (Abcam, ab23666), α-NDP52 (Abcam, ab68588), α-TAX1BP1 (Abcam, ab121812), α-NBR1 (Novus Biologicals, NBP1-71703), α-MAP1LC3B (NB100-2200), α-BNIP3 (Abcam, ab109362), α-FUNDC1 (ab74834), α-NIX (Abcam, ab8399), α-TOM20 (Abcam, ab56783, ab78547), α-Parkin (Novus Biologicals, NBP2-67017), α-HSP60 (Abcam, ab13532), α-SOD2 (Novus Biologicals, NB100-1992), α-PDHA1 (Abcam, ab110330), α-DRP1 (Abcam, ab184247), α-SNX9 (Abcam, ab181856), α-ECSIT (Novus Biologicals, NBP1-918858), α-LAMP1 (Santa Cruz Biotechnology, Sc-20011), MIRO1 (Novus, NBP1-89011), MIRO-2 (Novus, NBP1-88982), α-RAB7 (Abcam, ab50533), α-CATHEPSIN-D (Santa Cruz Biotechnology, Sc-10725), α-Ubiquitin (Abcam, ab134953), α-MCU (Cell Signaling Technology, D2Z3B), α-TUBULIN (Abcam, ab6046), α-ACTIN (Abcam, ab8226), α-GAPDH (Abcam, ab9485) All secondary antibodies used were from Invitrogen, conjugated with Alexa Flour dye: anti-mouse Alexa Fluor 405 (A31553), anti-mouse Alexa Fluor 488 (A28175), anti-mouse Alexa Fluor 568 (A11031), anti-mouse Alexa Fluor 647 (A21235), anti-rabbit Alexa Fluor 405 (A31556), anti-rabbit Alexa Fluor 488 (A11034), anti-rabbit Alexa Fluor 568 (A11011), anti-rabbit Alexa Fluor 647 (A21245).

Techniques: Infection, Membrane

Journal: BioMed Research International

Article Title: Cellular Levels of Oxidative Stress Affect the Response of Cervical Cancer Cells to Chemotherapeutic Agents

doi: 10.1155/2014/574659

Figure Lengend Snippet: Relative expression (SiHa versus CaSki) of genes involved in oxidative stress and antioxidant defense.

Article Snippet: Monoclonal α -NQO1, α -OXR1, and α - β -actin were obtained from Sigma-Aldrich, monoclonal α -SOD1, and α -GPX 1/2 from Santa Cruz Biotechnology, monoclonal α -SOD2 from BD Biosciences, and monoclonal α -PARP1 (Ab-2) from Millipore Corporation (Calbiochem). tert -Butyl hydroperoxide solution (tBHP), N -Acetyl-L-cysteine (NAC), cis -diammineplatinum(II) dichloride (cisplatin), doxorubicin hydrochloride (DOX), and DL-buthionine-(S-R)-sulfoximine (BSO) were purchased from Sigma-Aldrich.

Techniques: Expressing