Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Microbiota-gut-brain axis and neuroendocrine pathways underlie divergent mechanisms of intermittent and continuous theta-burst stimulation in autism spectrum disorder

doi: 10.1007/s00018-026-06096-2

Figure Lengend Snippet: The protein expression of PI3K, AKT, mTOR and their respective phosphorylated forms detected by Western blotting

Article Snippet: For immunofluorescence staining, free-floating hypothalamic sections were co-incubated with primary antibodies against somatostatin (SST) (1:100, rabbit, bs-37040R, Bioss) and growth hormone-releasing hormone (GHRH) (1:100, rabbit, bs-0205R, Bioss), while prefrontal cortical sections were incubated with primary antibodies targeting growth hormone (GH) (1:1000, rabbit, GB113303-100, Servicebio), phospho-AKT (1:100, rabbit, bs-0876R, Bioss), phospho-mTOR (1:100, rabbit, bs-3495R, Bioss), growth hormone receptor (GHR) (1:100, rabbit, bs-0654R, Bioss), and somatostatin receptor 2 (SSTR2) (1:100, rabbit, bs-10986R, Bioss).

Techniques: Expressing, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Microbiota-gut-brain axis and neuroendocrine pathways underlie divergent mechanisms of intermittent and continuous theta-burst stimulation in autism spectrum disorder

doi: 10.1007/s00018-026-06096-2

Figure Lengend Snippet: Immunofluorescence analysis of GHR and SSTR2 in the prefrontal cortex. A Proposed signaling pathway of GH in regulating the PI3K/Akt/mTOR axis (KEGG Entry: map04935). B Quantitative analysis of the expression levels of GHR and SSTR2. C Representative immunofluorescence images of GHR and SSTR2 (50.0×, Scale bar: 20 μm). The data are presented as mean ± SD ( n = 3). ### p < 0.001 vs. control group; *** p < 0.001 vs. VPA group

Article Snippet: For immunofluorescence staining, free-floating hypothalamic sections were co-incubated with primary antibodies against somatostatin (SST) (1:100, rabbit, bs-37040R, Bioss) and growth hormone-releasing hormone (GHRH) (1:100, rabbit, bs-0205R, Bioss), while prefrontal cortical sections were incubated with primary antibodies targeting growth hormone (GH) (1:1000, rabbit, GB113303-100, Servicebio), phospho-AKT (1:100, rabbit, bs-0876R, Bioss), phospho-mTOR (1:100, rabbit, bs-3495R, Bioss), growth hormone receptor (GHR) (1:100, rabbit, bs-0654R, Bioss), and somatostatin receptor 2 (SSTR2) (1:100, rabbit, bs-10986R, Bioss).

Techniques: Immunofluorescence, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Microbiota-gut-brain axis and neuroendocrine pathways underlie divergent mechanisms of intermittent and continuous theta-burst stimulation in autism spectrum disorder

doi: 10.1007/s00018-026-06096-2

Figure Lengend Snippet: Immunofluorescence analysis of GH and p-AKT/GH and p-mTOR co-localization in the prefrontal cortex. A Representative immunofluorescence images of GH and p-Akt (50.0×, Scale bar: 20 μm). B Quantification of the Pearson’s correlation coefficient (R value) for GH and p-Akt co-localization. C Representative immunofluorescence images of GH and p-mTOR (50.0×, Scale bar: 20 μm). D Quantification of the Pearson’s correlation coefficient (R value) for GH and p-mTOR co-localization from. The data are presented as mean ± SD ( n = 3). ### p < 0.001 vs. control group; *** p < 0.001 vs. VPA group

Article Snippet: For immunofluorescence staining, free-floating hypothalamic sections were co-incubated with primary antibodies against somatostatin (SST) (1:100, rabbit, bs-37040R, Bioss) and growth hormone-releasing hormone (GHRH) (1:100, rabbit, bs-0205R, Bioss), while prefrontal cortical sections were incubated with primary antibodies targeting growth hormone (GH) (1:1000, rabbit, GB113303-100, Servicebio), phospho-AKT (1:100, rabbit, bs-0876R, Bioss), phospho-mTOR (1:100, rabbit, bs-3495R, Bioss), growth hormone receptor (GHR) (1:100, rabbit, bs-0654R, Bioss), and somatostatin receptor 2 (SSTR2) (1:100, rabbit, bs-10986R, Bioss).

Techniques: Immunofluorescence, Control

Journal: Oncology Reports

Article Title: Saikosaponin D overcomes gemcitabine resistance in pancreatic cancer via AKT/mTOR pathway inhibition and synergistic induction of apoptosis and autophagy

doi: 10.3892/or.2025.9033

Figure Lengend Snippet: AKT/mTOR pathway inhibition by GEM and SSD combination therapy. (A) Immunocytochemistry staining of AKT/mTOR signaling proteins in MIA PaCa-2 and AsPC-1 cells treated with Ctrl, G0.25, S4 or the combination (G0.25 + S4; magnification, ×40; insets show phosphoprotein details). Scale bar, 50 µm. (B) Western blot analysis of p-AKT (60 kDa), AKT (56 kDa), p-mTOR (289 kDa), mTOR (289 kDa) and β-actin (42 kDa). Semi-quantification of phosphorylation ratios in (C) MIA PaCa-2 and (D) AsPC-1 cells. In the figure, GEM represents 0.25 µmol/l GEM and SSD represents 4 µmol/l SSD. Data are presented as the mean ± SD (n=5). Compared with GEM monotherapy, the G0.25 + S4 combination led to a greater reduction in both p-AKT/AKT and p-mTOR/mTOR ratios (both P adj <0.0001), with large effect sizes (η 2 =0.73 and 0.81, respectively). All P-values are Benjamini-Hochberg-adjusted. ****P<0.0001 for G0.25 + S4 vs. Ctrl; ## P<0.01, ### P<0.001 for G0.25 + S4 vs. G0.25. Statistical analysis was performed using one-way ANOVA with Tukey's post hoc test. Ctrl, control; GEM, gemcitabine; SSD, Saikosaponin D; p-, phosphorylated.

Article Snippet: The following antibodies were purchased from Proteintech Group, Inc.: Mouse monoclonal antibodies against mTOR (1:500; cat. no. 66888-1-Ig), phosphorylated (p-)mTOR (Ser2448) (1:2,000; cat. no. 67778-1-Ig), p-AKT (Ser473) (1:500; cat. no. 66444-1-Ig), caspase-3 (1:2,000; cat. no. 66470-2-Ig), cleaved caspase-3 (1:1,000; cat. no. 66470-2-Ig), Bax (1:500; cat. no. 60267-1-Ig), AKT (1:500; cat. no. 60203-2-Ig) and β-actin (1:2,000; cat. no. 66009-1-Ig); as well as rabbit antibodies, including polyclonal antibodies against Bcl-2 (1:1,000; cat. no. 12789-1-AP), Beclin 1 (1:1,000; cat. no. 11306-1-AP) and LC3 (1:1,000; cat. no. 14600-1-AP).

Techniques: Inhibition, Immunocytochemistry, Staining, Western Blot, Phospho-proteomics, Control

Journal: Oncology Reports

Article Title: Saikosaponin D overcomes gemcitabine resistance in pancreatic cancer via AKT/mTOR pathway inhibition and synergistic induction of apoptosis and autophagy

doi: 10.3892/or.2025.9033

Figure Lengend Snippet: Proposed mechanism of GEM and SSD combination therapy. Co-treatment with GEM and SSD synergistically inhibits the AKT/mTOR pathway, concurrently inducing apoptosis (via Bax/Bcl-2 modulation and caspase-3 activation) and autophagy (via Beclin 1 activation and LC3 conversion), thereby overcoming GEM resistance in pancreatic cancer cells. GEM, gemcitabine; P, phosphorylated; SSD, Saikosaponin D.

Article Snippet: The following antibodies were purchased from Proteintech Group, Inc.: Mouse monoclonal antibodies against mTOR (1:500; cat. no. 66888-1-Ig), phosphorylated (p-)mTOR (Ser2448) (1:2,000; cat. no. 67778-1-Ig), p-AKT (Ser473) (1:500; cat. no. 66444-1-Ig), caspase-3 (1:2,000; cat. no. 66470-2-Ig), cleaved caspase-3 (1:1,000; cat. no. 66470-2-Ig), Bax (1:500; cat. no. 60267-1-Ig), AKT (1:500; cat. no. 60203-2-Ig) and β-actin (1:2,000; cat. no. 66009-1-Ig); as well as rabbit antibodies, including polyclonal antibodies against Bcl-2 (1:1,000; cat. no. 12789-1-AP), Beclin 1 (1:1,000; cat. no. 11306-1-AP) and LC3 (1:1,000; cat. no. 14600-1-AP).

Techniques: Activation Assay

Journal: Research

Article Title: A Novel Autophagy Inhibitor p -Hydroxylcinnamaldehyde Suppresses Esophageal Squamous Cell Carcinoma by Targeting LDHA Phosphorylation-Mediated Metabolic Reprogramming

doi: 10.34133/research.1070

Figure Lengend Snippet: CMSP reduces AMPK/mTOR pathway activity in ESCC cells. (A) AMP, ATP, and ratio of AMP and ATP response to CMSP treatment in targeted metabolomics. (B) Representative Western blot of p-mTOR, mTOR, p-AMPK, AMPK, p-P70S6K, P70S6K, p-ULK1, and ULK1 following treatment of concentration-dependent CMSP for 36 h. (C) Representative Western blot of p-mTOR, p-AMPK, p-P70S6K, and p-ULK1 following treatment of CMSP (40 μg/ml) for different time. (D) Representative Western blot of p-mTOR, p-AMPK, p-P70S6K and p-ULK1 following treatment of CMSP (40 μg/ml) and rapamycin (20 nM). Data are shown as the mean ± SD ( n = 3); Student’s t test; ** P < 0.01 versus the control group.

Article Snippet: Antibodies against actin (20536-1-AP), LC3B (14600-1-AP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (10494-1-AP), p-mTOR (67778-1-Ig), mTOR (66888-1-Ig), AMPK (66536-1-Ig), P70S6K (14485-1-AP), and ULK1 (20986-1-AP) were purchased from Proteintech Group (Wuhan, China).

Techniques: Activity Assay, Western Blot, Concentration Assay, Control