Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: LM11A-31, a modulator of p75 neurotrophin receptor, suppresses HIV-1 replication and inflammatory response in macrophages.

doi: 10.3389/ebm.2024.10123

Figure Lengend Snippet: FIGURE 1 The effect of LM11A-31 on cytotoxicity, antioxidant enzymes, and autophagy marker in U937 macrophages. U937 macrophages were differentiated and then treated with DMSO, LM11A-31 (100 nM), DRV (5.5 μM), and LM11A-31 + DRV for 48 h. The cytotoxicity was measured using CyQUANT™LDH Cytotoxicity Assay Kit. (A) LDH measurement at 24 h. (B) LDH measurement at 48 h. (C) Quantification of protein level of catalase at 48 h. (D) Quantification of protein level of SOD1 at 48 h. (E) Quantification of protein level of Beclin-1 at 48 h. Values are expressed as mean ± SEM (n = 3), *p < 0.05.

Article Snippet: The HIV-1-uninfected U937 monocytic cell line and HIV-1infected U1 monocytic cell line were obtained from ATCC (Manassas, VA).

Techniques: Marker, CyQUANT Assay, LDH Cytotoxicity Assay

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: LM11A-31, a modulator of p75 neurotrophin receptor, suppresses HIV-1 replication and inflammatory response in macrophages.

doi: 10.3389/ebm.2024.10123

Figure Lengend Snippet: FIGURE 2 The effect of LM11A-31 on inflammatory response in U937 macrophages. U937 macrophages were differentiated and then treated with DMSO, LM11A- 31 (100 nM), DRV (5.5 μM), and LM11A-31 + DRV for 48 h. The cytokines and chemokines profile were measured using a customized human 9-Plex ProcartaPlex™multiplex immunoassay. (A) Proinflammatory, chemokines, and anti-inflammatory measurement at 24 h. (B) Proinflammatory, chemokines, and anti-inflammatory measurement at 48 h. Values are expressed as mean ± SEM (n = 3). *p < 0.05, **p < 0.01.

Article Snippet: The HIV-1-uninfected U937 monocytic cell line and HIV-1infected U1 monocytic cell line were obtained from ATCC (Manassas, VA).

Techniques: Multiplex Assay

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: LM11A-31, a modulator of p75 neurotrophin receptor, suppresses HIV-1 replication and inflammatory response in macrophages.

doi: 10.3389/ebm.2024.10123

Figure Lengend Snippet: FIGURE 6 The effect of p75NTR in LM11A-31-mediated viral replication in U1 macrophages. U1 macrophages were differentiated and then treated with DMSO, LM11A-31 (100 nM), DRV (5.5 μM), and LM11A-31 + DRV for 48 h. The protein expression was measured using a western blot. (A) Quantification of protein level of p75NTR in U1 and U937. (B, C) Quantification of protein level of p75NTR at 24 and 48 h. Values are expressed as mean ± SEM (n = 3–4), *p < 0.05.

Article Snippet: The HIV-1-uninfected U937 monocytic cell line and HIV-1infected U1 monocytic cell line were obtained from ATCC (Manassas, VA).

Techniques: Expressing, Western Blot

Journal: International journal of cancer

Article Title: Hematopoietic stem cell differentiation affects expression and function of MRP4 (ABCC4), a transport protein for signaling molecules and drugs.

doi: 10.1002/ijc.24207

Figure Lengend Snippet: FIGURE 6 – Expression and function of MRP4 in the U937 cell line differentiated in the presence of vitamin D3 and PMA. U937 cells were cultivated in growth medium (gray curve) or in the presence of the differentiating agents (black curve) vitamin D3 (a,c) or PMA (b,d) and stained with antibodies against CD11b (a,b) and CD14 (c,d). For negative control cells were incubated with normal mouse IgG1 instead of a specific primary antibody (transparent curve) or assessed without staining (broken transparent curve). Cells were cultivated in the pres- ence of 1 lM vitamin D3 (a, c) or 50 nM PMA (b, d). Results are rep- resentative for at least 3 independent differentiation experiments. (e) mRNA expression of MRP4, quantities were normalized to 18S rRNA (values represent mean 6 SD, n 5 3, *p < 0.05; one-way ANOVA). (f) For viability assays differentiated cells were incubated with either 100 lM 6-mercaptopurine (black columns) or 0.1 mM sodium hydrox- ide solution (white columns) and cell viability was assessed. Values rep- resent mean 6 SD (n 5 3; **p < 0.01; *p < 0.05; one-way ANOVA).

Article Snippet: 38–40 HL60 and U937 cell lines were purchased from DSMZ (Braunschweig, Germany), the M-07e cell line was kindly provided by Dr. Joachim Boos (Dept. of Pediatric Oncology, University of M€unster, Germany).

Techniques: Expressing, Staining, Negative Control, Incubation

Journal:

Article Title: The Cyclin A1-CDK2 Complex Regulates DNA Double-Strand Break Repair

doi: 10.1128/MCB.24.20.8917-8928.2004

Figure Lengend Snippet: Triple hybrid screening: interaction of cyclin A1 with Ku70/Ku80. (A) A yeast triple hybrid screen was performed with a Gal4-binding domain-cyclin A1 fusion protein as the bait, while human CDK2 was conditionally expressed. Screening was performed with a human testis cDNA library. (B) Two of the identified interacting clones contained a fragment of the human Ku70 cDNA. (C) Interaction of cyclin A1 and Ku70 in vitro. Baculovirus-expressed cyclin A1 or cyclin A1/CDK2 was incubated with GST, GST-Ku70, or GST-Ku80. After GST pulldown, binding was detected with anti-cyclin A1 antibody or anti-CDK2 antibody. (D) Deletion mutants of Ku70 were tested for their interaction with cyclin A1 in GST pulldown assays. Cyclin A1 binding was detected with anti-cyclin A1 Western blotting. The C terminus of Ku70 was required for the interaction with cyclin A1. (E) Interaction of cyclin A1 and Ku70 in vivo. Immunoprecipitations were performed with either anti-cyclin A1 or control immunoglobulin G. Ku70 coimmunoprecipitated with cyclin A1 in U937 cell lysate. (F) GST-Ku70, GST, or GST-Ku80 was incubated with baculovirus-expressed cyclin A1, CDK2, or cyclin A1/CDK2 in the presence of [γ-32P]ATP. Lysate from wild-type baculovirus-infected insect cells was used as a control. Following SDS-PAGE, the gel was autoradiographed. GST-Ku70 protein was clearly phosphorylated by cyclin A1/CDK2, whereas GST-Ku80 was not. The nonspecific higher band appeared in all three GST preparations (GST, GST-Ku70, and GST-Ku80).

Article Snippet: U937 cells were lysed in radioimmunoprecipitation (RIPA) buffer; 300 μg of cell lysate was used in the immunoprecipitation assays with 3 μg of primary antibody in 500 μl of RIPA buffer for 2 h at 4°C; 50 μl of a 50% slurry of protein A/G-Plus agarose (Santa Cruz Biotechnology) was added for another 1 h, and beads were washed three times with RIPA buffer.

Techniques: Binding Assay, cDNA Library Assay, Clone Assay, In Vitro, Incubation, Western Blot, In Vivo, Infection, SDS Page