Journal: Cell reports

Article Title: Altered GM1 catabolism affects NMDAR-mediated Ca 2+ signaling at ER-PM junctions and increases synaptic spine formation in a GM1-gangliosidosis model

doi: 10.1016/j.celrep.2024.114117

Figure Lengend Snippet: (A) Immunoblot analysis of cell fractions from 6-month-old WT and β-Gal − / − mice. Markers for the ER (Calnexin), PM (N-cadherin), and ER-PM junctions (ORAI1, STIM1, STIM2, VAPA, and VAPB) were enriched in their respective fractions. Immunoblots using HRP-conjugated cholera toxin B subunit (CTX-B) show high GM1 levels in β-Gal − / − fractions. (B) Representative HPTLC plate showing GM1 levels in the ER, PM, and ER-PM junctions isolated from 6-month-old WT and β-Gal − / − mice. STD, standard. Note: to detect GM1 in WT samples, the sample volume loaded was 3× that of the β-Gal − / − samples. (C) Quantification of GM1 levels from HPTLC plates performed in (B). n = 8. Values are expressed as median ± quartiles. Statistical analysis was performed using the Student’s t test; *** p < 0.001, **** p < 0.0001. (D) Representative HPTLC plate showing GM1 levels in ER-PM junctions isolated from 1-, 3-, and 6-month-old WT and β-Gal − / − mice. To detect GM1 in WT samples, the sample volume loaded was 3× that of the β-Gal − / − samples. (E) Quantification of GM1 levels from HPTLC plates performed in (D). n = 4. Values are expressed as median ± quartiles. Statistical analysis was performed using the Student’s t test with Welch’s correction; ns, not significant; *** p < 0.001, **** p < 0.0001.

Article Snippet: Rabbit anti-STIM2 antibody , Cell Signaling , Cat #: 4917; RRID: AB_2198021.

Techniques: Western Blot, High Performance Thin Layer Chromatography, Isolation

Journal: Cell reports

Article Title: Altered GM1 catabolism affects NMDAR-mediated Ca 2+ signaling at ER-PM junctions and increases synaptic spine formation in a GM1-gangliosidosis model

doi: 10.1016/j.celrep.2024.114117

Figure Lengend Snippet:

Article Snippet: Rabbit anti-STIM2 antibody , Cell Signaling , Cat #: 4917; RRID: AB_2198021.

Techniques: Virus, Recombinant, Isolation, Magnetic Beads, Plasmid Preparation, Software

Journal: Molecular oncology

Article Title: Postbiotics of Lacticaseibacillus paracasei CECT 9610 and Lactiplantibacillus plantarum CECT 9608 attenuates store-operated calcium entry and FAK phosphorylation in colorectal cancer cells.

doi: 10.1002/1878-0261.13629

Figure Lengend Snippet: Fig. 1. Expression of Orai and stromal interaction molecule (STIM) proteins and Store-operated Ca2+ entry (SOCE) is enhanced in the colorectal adenocarcinoma cell lines HT-29 and Caco-2. (A–E) NCM460, HT-29 and Caco-2 cells were lysed and the whole cell lysates were analyzed by western blotting using anti-Orai1 (A), anti-Orai2 (B), anti-Orai3 (C), anti-STIM1 (D) or anti-STIM2 (E) antibody. Molecular masses indicated on the right were determined using molecular-mass markers run in the same gel. Membranes were probed with anti-b-actin antibody for protein loading control. These results are representative of four separate experiments. Quantification of protein expression in NCM460 (n = 4), HT-29 (n = 4) and Caco-2 (n = 4) cells normalized to the b-actin expression is depicted in the bar graph. Data are represented as mean standard error of the mean (SEM) and were statistically analyzed using Kruskal–Wallis test combined with Dunn’s post hoc test. **P < 0.01 and ****P < 0.0001. (F) Representative Ca2+ mobilization in response to 2 lM thapsigargin (TG) measured using fura-2 in NCM460 (n = 3 [62 cells]), HT-29 (n = 3 [69 cells]) and Caco-2 (n = 3 [36 cells]) cells. Cells were superfused with a Ca2+-free Hepes Buffer Saline (HBS) (100 lM ethylene glycol-bis(2-aminoethylether)-N,N,N0,N0-tetraacetic acid (EGTA) added) and stimulated with 2 lM TG, followed by re-addition of CaCl2 (1.8 mM) to estimate Ca2+ influx. (G, H) Quantification of TG-evoked Ca2+ release from the intracellular stores and entry in NCM460 (n = 3 [62 cells]), HT-29 (n = 3 [69 cells]) and Caco-2 (n = 3 [36 cells]) cells is shown in the scatter plots. Data in bar graphs are represented as mean SEM and were statistically analyzed using Kruskal–Wallis test combined with Dunn’s post hoc test. ****P < 0.0001. a.u., arbitrary units; AUC, area under the curve.

Article Snippet: Rabbit polyclonal anti-STIM2 antibody (catalog number 4917S) was from Cell Signaling Technology (Leiden, the Netherlands).

Techniques: Expressing, Western Blot, Control, Saline