Journal: The Journal of biological chemistry

Article Title: Lactate promotes neuronal differentiation of SH-SY5Y cells by lactate-responsive gene sets through NDRG3-dependent and -independent manners.

doi: 10.1016/j.jbc.2023.104802

Figure Lengend Snippet: Figure 11. TEAD1 and ELF4 differently affect lactate-NDRG3-regulated neuronal maker gene expressions in SH-SY5Y cells. A–C, the protein expression levels of NF-H, RUNX1T1 and SYT4 in siNDRG3 (A), siTEAD1 (B), and siELF4 (C) or scrambled siRNA transfected SH-SY5Y cells (n = 4). Relative protein levels were normalized to the levels of RPL13A and α-actinin in the same samples and presented as fold change to control. All data are reported as means ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control. Statistical significance was determined by unpaired two-tailed t test. Since these protein expression levels were analyzed by the same lysate in Figure 8C, the blots of RPL13a, α-actinin, and NDRG3 were the same and used for the normalization. D, a schematic illustration of lactate-induced neuronal differentiation mechanism through NDRG3-dependent and -independent signaling axis.

Article Snippet: The membranes were blocked in 5% fat-free milk in TBST (20 mM Tris-HCl (pH 7.6), 0.15 M sodium chloride, and 0.1% Tween 20) for 1 h, washed three times with TBST, and incubated with primary antibodies that probe for NSE (9536, CST), ID2 (3431, CST), MAP2 (4542, CST), NF-H (ab4680, abcam), RPL13A (2765, CST), α-actinin (6487, CST), Cyclophilin B (43603, CST), β-actin (8457, CST), NDRG3 (ab133715, abcam), MCT1 (20139-1-AP, Proteintech), MCT2 (ab224627, abcam), MCT4 (ab74109, abcam), TEAD1 (12292, CST), ELF4 (sc-390689, Santacruz), EEF1A2 (GTX102326, GeneTex), HES7 (AP9712a, Abcepta), TLE2 (GTX106107, GeneTex), RUNX1T1 (15494-1-AP, Proteintech) and SYT4 (12642-1-AP, Proteintech) in TBST for overnight.

Techniques: Expressing, Transfection, Control, Two Tailed Test

Journal: Leukemia Research

Article Title: Directed Therapy for Patients with Myelodysplastic Syndromes (MDS) by Suppression of Cyclin D1 with ON 01910.Na

doi: 10.1016/j.leukres.2012.04.002

Figure Lengend Snippet: Decreasing levels of cyclin D1 in peripheral blood CD34+ cells following administration of ON 1910.Na Cyclin D1 was measured by flow cytometry as described in Materials and Methods in peripheral bood CD34+ cells of patients receiving ON1910. Gating strategy is seen at far left: live CD33+ cells were discriminated from dead cells by a CD33 versus BiViD gate. Mononuclear cells were identified by a forward scatter-area (FSC-A) versus side scatter-area (SSC-A) plot. Viable CD33+ cells were then dichotomized on the basis of maturation by a CD13+ versus CD34+ bivariate plot. Cyclin-D1 positives were then identified by a CD33 versus cyclin-D plot, with positive events for cyclin-D1 defined on the basis of a fluorescence-minus-one (FMO) control. Percentages of CD34+ cells expressing cyclin D1 from three patients are seen in the right panel.

Article Snippet: Cells were stained intracellularly with cyclin-D mABs (BD Pharmingen) and analyzed using a LSR-II or LSR Fortessa flow cytometer (BD).

Techniques: Flow Cytometry, Fluorescence, Expressing