Journal: Autophagy

Article Title: Trophoblast aging driven by IL33 deficiency elevates recurrent pregnancy loss risk through SNAP29 lactylation-mediated autophagy impairment

doi: 10.1080/15548627.2026.2659946

Figure Lengend Snippet: Senescence-induced glycolysis and lactate accumulation impair autophagy in trophoblasts. ( A ) Heatmap showed the relative expression changes of seven proteins in the glycolysis/gluconeogenesis pathway. ( B ) Glucose and lactate concentrations in the supernatant of senescent HTR-8/SVneo cells treated with or without IL33 (10 ng/mL). ( C and D ) Western blot for SQSTM1 or LC3B-II:I in normal and senescent HTR-8/SVneo cells treated with different concentration of lactate (0, 10, 20 mM for 48 h). Relative expression levels of proteins were standardized using internal reference ACTB. ( E ) The expression of lactylation level was analyzed by immunohistochemistry staining in normal and URPL villi ( n = 6 per group). Scale bar: 200 μm. ( F ) The expression of lactylation level was analyzed by immunohistochemistry staining in wt and il33 −/− mice ( n = 6 per group). Scale bar: 500 μm. ( G ) Western blot for lactylation level in HTR-8/SVneo cell senescence models treated with or without IL33 (10 ng/mL). Relative expression levels of proteins were standardized using internal reference histone H3. ( H ) Western blot for lactylation level in normal and senescent HTR-8/SVneo cells treated with different concentration of lactate (0, 10, 20 mM for 48 h). Relative expression levels of proteins were standardized using internal reference histone H3. ( I ) Western blot for lactylation and acetylation level in trophoblast cells treated with tacedinaline (5 μM, 48 h) or TMP195 (5 μM, 48 h). Relative expression levels of proteins were standardized using internal reference histone H3. ( J ) The expression of SQSTM1 was detected by immunofluorescence in HTR-8/SVneo cells treated with tacedinaline (5 μM, 48 h) or TMP195 (5 μM, 48 h). Scale bar: 10 μm. ( K and L ) The level of autophagy was detected by the DAL/DAP fluorescence probes in HTR-8/SVneo cells treated with tacedinaline (5 μM, 48 h) or TMP195 (5 μM, 48 h). Scale bar: 50 μm. Immunofluorescence staining results was quantified with average intensity. All data were generated using ImageJ. Statistical data were presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significant difference , using two-tailed Student’s t-test (e, F) and one-way ANOVA test (b, d, L).

Article Snippet: IL33 (10 ng/mL; PeproTech, 200–33), rapamycin (2 μM, 24 h; Sigma-Aldrich, V900930), lactate (10 or 20 mM, 48 h; Sigma-Aldrich, L6661), tacedinaline (5 μM, 48 h; MedChem Express, HY-50934), TMP195 (5 μM, 48 h; MedChem Express, HY-18361), 2-deoxy-D-glucose (2-DG; 5 or 10 mM, 48 h; MedChem Expres, HY-13966s), Cycloheximide (CHX, 30 μg/mL, 0–8 h; MedChem Express, HY-12320), MG132 (10 μM, 9 h; MedChem Express, HY-13259), bafilomycin A 1 (BafA1, 100 nM, 9 h; MedChem Express, HY-100558) were used.

Techniques: Expressing, Western Blot, Concentration Assay, Immunohistochemistry, Staining, Immunofluorescence, Fluorescence, Generated, Two Tailed Test

Journal: Biological Research

Article Title: Inhibition of phospholipase D2 augments histone deacetylase inhibitor-induced cell death in breast cancer cells

doi: 10.1186/s40659-020-00294-3

Figure Lengend Snippet: PLD2 inhibition sensitizes SAHA-induced apoptotic cell death. a MDA-MB-231 cells were transfected with siRNA of PLD2 and then treated with vehicle or SAHA (5 μM) for 36 h. Apoptosis was examined via flow cytometry and presented as the percentage of cells labeled with annexin V-PE. b MDA-MB-231 cells were stained with propidium iodide (1 μg/mL) and the population of subG1 apoptotic cells was determined using FACScan flow cytometry. c MDA-MB231 cells were treated with T247 (3 μM), TMP195 (3 μM), and PLD2 inhibitor (10 μM) for 36 h, and caspase-3 activity was examined. d MDA-MB231 were treated with SAHA (5 μM), rapamycin (50 nM), temozolomide (50 μM), and PLD2 inhibitor (10 μM) for 36 h, and caspase-3 activity was examined. e The cells were treated with SAHA, PLD2 inhibitor, or PA (50 μM) for 36 h. Apoptosis using annexin V staining was examined. f MDA-MB-231 cells were treated with the indicated drugs for 36 h, and caspase-3 activity and g expression of the indicated proteins were examined. The intensity of the indicated bands was normalized to the intensity of their respective α-tubulin bands and quantified against each other. Results are representative of at least four independent experiments and shown as the mean ± SEM. ** p < 0.001

Article Snippet: T247 and TMP195 were from MedKoo Biosciences (Morrisville, NC, USA).

Techniques: Inhibition, Transfection, Flow Cytometry, Labeling, Staining, Activity Assay, Expressing