Journal: iScience

Article Title: AP39 through AMPK-ULK1-FUNDC1 pathway regulates mitophagy, inhibits pyroptosis, and improves doxorubicin-induced myocardial fibrosis

doi: 10.1016/j.isci.2024.109321

Figure Lengend Snippet: Effect of targeted mitochondrial H 2 S on the structure and fibrosis of DOX-treated rat myocardial tissue (A) HE staining of rat myocardial tissue, pink is the cytoplasm, blue part is the nucleus (Scale bar: 20 μm). (B) Immunohistochemical staining results of collagen III of rat myocardial tissue, yellow part is collagen III positive (Scale bar: 20 μm). (C) Masson staining of rat myocardial tissue, the blue part is collagen fiber (Scale bar: 20 μm). (D) Calculated results of the collagen volume fraction of rat myocardial tissue in each group. (E) Calculated results of collagen III immunohistochemistry of rat myocardial tissue in each group. (F) Changes in the mRNA expression levels of Col2a1 . (G–L) Changes in collagen III, α-SMA, MMP13, MMP8, and TIMP1 protein expression levels in rat myocardial tissue and corresponding semiquantitative analysis results. Data were represented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: The proteins were wet transferred to PVDF membranes and blocked with a 5% fat-free milk-blocking solution for 1 h. CSE (12217-1-AP), α-SMA (14395-1-AP), collagen III (22734-1-AP), MMP8 (17874-1-AP), MMP13 (18165-1-AP), TIMP1 (6644- 1-AP), caspase1 (22915-1-AP), IL-1β (16806-1-AP), beclin1 (11306-1-AP), LC3 (18725-1-AP), P62 (18420-1-AP), AMPK (10929-2-AP), ULK1 (20986-1-AP), and GAPDH (60004-1-IG) were purchased from Proteintech (Chicago, USA). p -AMPK (2535), GSDMD (39754), and NLRP3 (15101) were purchased from CST (Cell Signaling Technology, Boston, MA, USA). p -ULK1 (ab133747) primary antibody was purchased from Abcam (Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Immunohistochemistry, Expressing

Journal: iScience

Article Title: AP39 through AMPK-ULK1-FUNDC1 pathway regulates mitophagy, inhibits pyroptosis, and improves doxorubicin-induced myocardial fibrosis

doi: 10.1016/j.isci.2024.109321

Figure Lengend Snippet:

Article Snippet: The proteins were wet transferred to PVDF membranes and blocked with a 5% fat-free milk-blocking solution for 1 h. CSE (12217-1-AP), α-SMA (14395-1-AP), collagen III (22734-1-AP), MMP8 (17874-1-AP), MMP13 (18165-1-AP), TIMP1 (6644- 1-AP), caspase1 (22915-1-AP), IL-1β (16806-1-AP), beclin1 (11306-1-AP), LC3 (18725-1-AP), P62 (18420-1-AP), AMPK (10929-2-AP), ULK1 (20986-1-AP), and GAPDH (60004-1-IG) were purchased from Proteintech (Chicago, USA). p -AMPK (2535), GSDMD (39754), and NLRP3 (15101) were purchased from CST (Cell Signaling Technology, Boston, MA, USA). p -ULK1 (ab133747) primary antibody was purchased from Abcam (Cambridge, UK).

Techniques: Recombinant, ROS Assay, Software, Imaging

Journal:

Article Title: OX40 Stimulation by gp34/OX40 Ligand Enhances Productive Human Immunodeficiency Virus Type 1 Infection

doi: 10.1128/JVI.75.15.6748-6757.2001

Figure Lengend Snippet: Enhanced expression of HIV-1 by OX40 stimulation from HIV-1 chronically infected ACH-2/OX40 cells. (A) Five groups of ACH-2/OX40 cells (cell groups 3, 4, 5, 6, and 10), four groups ACH-2/mock cells (cell groups 2, 7, 8, and 10), and the parental ACH-2 cell line were stimulated by PFA-fixed SV-T2/gp34 cells or PFA-fixed SV-T2/mock cells at a cell ratio of 2:1 with TNF-α (2 ng/ml) or TNF-β (10 ng/ml). After 3 days, HIV-1 production was determined quantitatively by p24 assay using the culture supernatants. (B) PFA-fixed SV-T2/gp34 cells and PFA-fixed SV-T2/mock cells were preincubated with or without anti-gp34 neutralizing MAb (5A8) or control MAb (Cont.) at 5, 1, 0.2, or 0.04 μg/ml for 1 h at 37°C and then were added to ACH-2/OX40 cells. Culture supernatants were harvested 3 days after coculture. (C) Kinetics of HIV-1 expression after stimulation with gp34 in ACH-2/OX40 cells. Supernatants were harvested at each time point, i.e., at 0, 2, 4, 8, 12, 18, 24, 48, 72, 96, and 120 h, and then tested for p24 titers. All results are the means of values obtained in triplicate. A representative result from two independent experiments is shown.

Article Snippet: Other MAbs used were fluorescein isothiocyanate (FITC)-conjugated anti-gp34 MAb (TAG-34) ( 40 ), Cy5-conjugated negative control mouse IgG1 (TAXY-8) ( 42 ), FITC-conjugated negative control mouse IgG1 (DAK-GO1; Dako, Copenhagen, Denmark), anti-human OX40 MAb (Ber Act 35; Ancell, Bayport, Minn.), anti-myc MAb (Invitrogen), anti-Fas MAb (CH-11; MBL, Nagoya, Japan), gp34-neutralizing MAb (5A8) ( 16 , 44 ), TNF-α-neutralizing MAb (1825.121; R&D Systems), TNF-β-neutralizing MAb (5802.21; R&D Systems), FasL-neutralizing MAb (NOK2; PharMingen, San Diego, Calif.), and negative control IgG1 (MET-3) ( 41 ).

Techniques: Expressing, Infection

Journal:

Article Title: OX40 Stimulation by gp34/OX40 Ligand Enhances Productive Human Immunodeficiency Virus Type 1 Infection

doi: 10.1128/JVI.75.15.6748-6757.2001

Figure Lengend Snippet: Autocrine secretion of TNF-α and TNF-β does not account for enhanced HIV-1 expression in ACH-2/OX40 cells stimulated with gp34. ACH-2/OX40#10 cells were cocultured with PFA-fixed SV-T2/gp34 cells or PFA-fixed SV-T2/mock cells at a cell ratio of 2:1 or were cultured in the presence of TNF-α (2 ng/ml) or TNF-β (10 ng/ml) after preincubation with or without anti-human TNF-α neutralizing MAb (50 μg/ml), anti-human TNF-β neutralizing MAb (5 μg/ml), or control MAb (50 μg/ml) for 1 h at 37°C. Culture supernatants were harvested after 3 days, and then the HIV-1 p24 level was measured. The results are the means of values obtained in triplicate. Representative results are shown from three independent experiments.

Article Snippet: Other MAbs used were fluorescein isothiocyanate (FITC)-conjugated anti-gp34 MAb (TAG-34) ( 40 ), Cy5-conjugated negative control mouse IgG1 (TAXY-8) ( 42 ), FITC-conjugated negative control mouse IgG1 (DAK-GO1; Dako, Copenhagen, Denmark), anti-human OX40 MAb (Ber Act 35; Ancell, Bayport, Minn.), anti-myc MAb (Invitrogen), anti-Fas MAb (CH-11; MBL, Nagoya, Japan), gp34-neutralizing MAb (5A8) ( 16 , 44 ), TNF-α-neutralizing MAb (1825.121; R&D Systems), TNF-β-neutralizing MAb (5802.21; R&D Systems), FasL-neutralizing MAb (NOK2; PharMingen, San Diego, Calif.), and negative control IgG1 (MET-3) ( 41 ).

Techniques: Expressing, Cell Culture