Journal: Journal of Alzheimer's Disease Reports

Article Title: Social interaction mitigates cognitive impairments and tau pathology in socially isolated aged mice

doi: 10.1177/25424823251394408

Figure Lengend Snippet: Co-housing alleviates synaptic damage in aged mice following social isolation. (A) Immunoblotting and (B) quantification of PSD93, PSD95, and synapsin-1 in hippocampus of two group mice (n = 4 per group, *** p < 0.001, two-tailed unpaired t-test). (C) Representative immunofluorescence images of synapsin-1 in DG and CA1 (green: synapsin-1, scale bar = 30 μm). Data expressed as mean ± SEM.

Article Snippet: Anti-PSD93 , Total PSD93 , mAb , Rabbit , Cell Signaling (19046S).

Techniques: Isolation, Western Blot, Two Tailed Test, Immunofluorescence

Journal: Cell death and differentiation

Article Title: Anaplastic Lymphoma Kinase signaling stabilizes SLC3A2 expression via MARCH11 to promote neuroblastoma cell growth.

doi: 10.1038/s41418-024-01319-0

Figure Lengend Snippet: Fig. 3 Knockdown of SLC3A2 suppresses NB cell growth and increases retinoic acid-induced neuronal differentiation. A–D CLB-BAR, CLB- GE, and CLB-GAR cells were treated with either control siRNA or SLC3A2-targeting siRNAs. A Cell growth was monitored by Incucyte every 6 h for 72-120 h (n = 3, mean ± s.e.m.). B Lysates collected at 72 h after siRNA transfection were immunoblotted with anti-SLC3A2, pAKT(S473), AKT, pERK (T202/Y204), ERK, PARP (asterisk (*) marked cleaved PARP), γH2A.X, α-tubulin and GAPDH. GAPDH was employed as immunoblotting loading control. Quantification of cleaved PARP (PARP*) and γH2A.X were performed with ImageJ (n = 3), shown in Supplementary Fig. 3. C Cell cycle analysis performed by NucleoCounter NC-3000 after 72 h (n = 3, mean ± s.e.m.). D Representative images showing differentiation morphology of CLB-GE cells 72 h post siRNA transfection. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells following siRNA treatment (n = 5, mean ± s.e.m). E, F SK-N-BE(2) cells treated with either SLC3A2-targeting or control siRNAs for 24 h, followed by retinoic acid (RA, 5 µM) for 48 h. E Representative images showing differentiation morphology of SK-N-BE(2) cells. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells (n = 6, mean ± s.e.m.). F Immunoblotting for neuronal differentiation markers (RET, DLG2) in SK-N-BE(2) cells (n = 3). GAPDH was employed as immunoblotting loading control. Immunoblot experiments were performed in biologically independent triplicates. Quantification was performed with ImageJ and analyzed by Student t test (unpaired, two- tailed). p values are indicated (****P < 0.0001, ***0.0001 < P < 0.001, **0.001 < P < 0.01, *0.01 < P < 0.05, ns P ≥0.05); exact P-values are shown in Supplementary Fig. 3 and Supplementary Table 1.

Article Snippet: Primary antibodies against human SLC3A2 (#81977, was employed for immunoblotting, 1:2000; #13180, was used for immunoprecipitation), ALK (#3633, 1:2000), pALK (Y1278, #6941, 1:1000), ubiquitin (#3936, 1:1000), phospho-Tyrosine (p-Y-1000, #8954, 1:1000), pAKT (S473,#4060, 1:4000), AKT (#9272), pERK1/2 (Y204/T202, #4377, 1;2000), PARP (#9542, 1:1000), γH2A.X (Ser139, #9718, 1:1000), RET (#14556, 1:2000), DLG2 (#19046, 1:1000), SLC1A5 (ASCT2, #8057, 1:2000), SLC7A5 (LAT1, #32683, 1:1000), SLC7A11 (xCT, #12691, 1:1000), SLC38A1 (SNAT1, #36057, 1:2000), β-actin (#4970, 1:10000), GAPDH (#5174, 1:10,000) and α-tubulin (#2125, 1:10,000) were obtained from Cell Signaling Technology.

Techniques: Knockdown, Control, Transfection, Western Blot, Cell Cycle Assay, Two Tailed Test

Journal: Cell death and differentiation

Article Title: Anaplastic Lymphoma Kinase signaling stabilizes SLC3A2 expression via MARCH11 to promote neuroblastoma cell growth.

doi: 10.1038/s41418-024-01319-0

Figure Lengend Snippet: Fig. 5 Combined lorlatinib and AMXT-1501 treatment synergistically inhibits cell growth in ALK-driven NB and increases retinoic acid- induced neuronal differentiation. A IC50s of AMXT-1501 in CLB-BAR, CLB-GE, SK-N-BE(2) and SH-SY5Y cells after 5 d treatment (n = 3, mean ± s.e.m.). B CLB-BAR and CLB-GE cells were treated with lorlatinib and AMXT-1501 at the indicated concentrations and cell growth monitored by Incucyte every 6 h over a total of 120 h (n = 3, mean ± s.e.m.). The combination index value was calculated based on Chou- Talalay method using CompuSyn software [70]. C Immunoblot analysis of ALK signaling and apoptosis markers in CLB-GE cells treated with AMXT-1501 (0 or 8 µM) and lorlatinib (0 or 16 nM) as indicated for 24 h (n = 3). Lysates were immunoblotted with anti-ALK, pY1278-ALK, pAKT(S473), AKT, pERK (T202/Y204), ERK, PARP (asterisk (*) marked cleaved PARP), γH2A.X, α-tubulin and SLC3A2 and β-actin. β-actin was employed as immunoblotting loading control. Representative images of differentiation morphology of SK-N-BE(2) (D) and SH-SY5Y (E) cells after AMXT-1501 (0 or 8 µM) and RA (0 or 5 µM) treatment as indicated for 48 h or 24 h. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells (n = 3, mean ± s.e.m.). F Immunoblot analysis of neuronal differentiation markers (RET and DLG2) in SK-N-BE(2) and SH- SY5Y cells treated with AMXT-1501 (0 or 8 µM) and RA (0 or 5 µM) for 48 h or 24 h (n = 3). β-actin was employed as immunoblotting loading control. Immunoblot experiments (C and F) were performed in biologically independent triplicates. Quantifications were performed with ImageJ and analyzed by Student t-test (unpaired, two-tailed). p values are indicated (****P < 0.0001, ***0.0001 < P < 0.001, **0.001 < P < 0.01, *0.01 < P < 0.05, ns P ≥0.05); exact P values are shown in Supplementary Fig. 4 and Supplementary Table 1.

Article Snippet: Primary antibodies against human SLC3A2 (#81977, was employed for immunoblotting, 1:2000; #13180, was used for immunoprecipitation), ALK (#3633, 1:2000), pALK (Y1278, #6941, 1:1000), ubiquitin (#3936, 1:1000), phospho-Tyrosine (p-Y-1000, #8954, 1:1000), pAKT (S473,#4060, 1:4000), AKT (#9272), pERK1/2 (Y204/T202, #4377, 1;2000), PARP (#9542, 1:1000), γH2A.X (Ser139, #9718, 1:1000), RET (#14556, 1:2000), DLG2 (#19046, 1:1000), SLC1A5 (ASCT2, #8057, 1:2000), SLC7A5 (LAT1, #32683, 1:1000), SLC7A11 (xCT, #12691, 1:1000), SLC38A1 (SNAT1, #36057, 1:2000), β-actin (#4970, 1:10000), GAPDH (#5174, 1:10,000) and α-tubulin (#2125, 1:10,000) were obtained from Cell Signaling Technology.

Techniques: Software, Western Blot, Control, Two Tailed Test