Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Mast Cells Inhibit Stem Cell-driven Epithelial Repair in Inflammatory Bowel Disease via Suppressing Wnt/lrp6/β-Catenin Signaling Pathway

doi: 10.1016/j.jcmgh.2026.101741

Figure Lengend Snippet: The effect of CPA inhibitor on ISCs-mediated epithelial regeneration in DSS-induced colitis. ( A ) Schematic of the in vivo experimental workflow. ( B ) Body weight loss: Body weight was recorded daily for each group. ( C ) DAI score: DAI was evaluated daily based on a standard scoring system (combining stool consistency, bleeding, and weight loss) to assess the severity of colitis. ( D and E ) Colon length measurement: Representative images of colon tissues ( D ) and quantitative analysis of colon length ( E ). ( F and G ) H&E staining of colon tissue: H&E-stained sections were observed under a light microscope to visualize pathological changes ( F ), and histological analysis was performed to quantify epithelial injury score ( G ). ( H and I ) Expression levels of ISC-related ( H ) and stemness-related ( I ) genes: qRT-PCR was used to measure the mRNA expression of ISC markers (Lgr5, Lrig1, Tert) and stemness-associated genes (Dek). ( J ) Expression levels of genes related to Wnt signaling pathways. ( K and L ) Representative images showing IF staining of total β-catenin ( K ) and active β-catenin ( L ) in colon (n = 3/group). ( M ) WB analysis of LRP6 and LGR5 proteins: WB was performed with β-actin as the loading control. and quantification analysis was performed to compare the differences. Statistical significance: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: To further examine whether MCs suppress ISC-mediated epithelial regeneration via CPA3, a CPA inhibitor (10 μM, MCE, HY-70005) was introduced into the co-culture system of activated BMMCs and organoids.

Techniques: In Vivo, Staining, Light Microscopy, Expressing, Quantitative RT-PCR, Protein-Protein interactions, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Mast Cells Inhibit Stem Cell-driven Epithelial Repair in Inflammatory Bowel Disease via Suppressing Wnt/lrp6/β-Catenin Signaling Pathway

doi: 10.1016/j.jcmgh.2026.101741

Figure Lengend Snippet: The effect of CPA inhibitor on MC-induced inhibition of ISC-mediated epithelial regeneration. Four groups were set: Control (organoids + Substance P + DMSO), CPA inhibitor (organoids + Substance P + CPA inhibitor), BMMC (organoids + activated BMMCs + DMSO), BMMC + CPA inhibitor (organoids + activated BMMCs + CPA inhibitor). All co-cultures lasted 3 days before analysis. ( A and B ) Organoid growth phenotypes after CPA inhibitor intervention. ( A ) Bright-field microscopy images showing organoid morphology in the 4 groups. ( B ) Quantitative analysis of organoid size, bud number, and density. ( C–F ) Gene expression analysis via qRT-PCR. Relative mRNA levels of ISC marker genes ( C ), stemness-associated genes ( D ), Wnt pathway genes ( E ), and apoptosis-related genes ( F ). ( G and H ) Representative images showing IF staining of total β-catenin ( G ) and active β-catenin ( H ) in colon (n = 4–6 organoids/group). Statistical significance: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: To further examine whether MCs suppress ISC-mediated epithelial regeneration via CPA3, a CPA inhibitor (10 μM, MCE, HY-70005) was introduced into the co-culture system of activated BMMCs and organoids.

Techniques: Inhibition, Control, Microscopy, Gene Expression, Quantitative RT-PCR, Marker, Staining

Journal: International Journal of Molecular Sciences

Article Title: Metformin Enhances Nomegestrol Acetate Suppressing Growth of Endometrial Cancer Cells and May Correlate to Downregulating mTOR Activity In Vitro and In Vivo

doi: 10.3390/ijms20133308

Figure Lengend Snippet: Nomegestrol acetate (NOMAC) simultaneously inhibited the growth of both RL95-2 and HEC-1A cells. ( A ) The structures of nomegestrol acetate (NOMAC), medroxyprogesterone acetate (MPA), levonorgestrel (LNG), or cyproterone acetate (CPA). ( B , C and D ) Inhibitory effect of four progestins on the viability of RL95-2, HEC-1A, and KLE cells. Cells were treated with NOMAC, MPA, LNG, or CPA at the concentration of 1, 3, 10, 30, and 100 μM for 48 h, respectively. Experiments were performed in triplicate each time and independently repeated three times with different passages of cells. The data were expressed as mean ± SEM. * p < 0.05, ** p < 0.01 vs. NOMAC treated cells.

Article Snippet: Levonorgestrel (LNG) were obtained from Beijing Zizhu Pharmaceutical Co. Ltd. (Beijing, China) and cyproterone (CPA) was purchased from Medchemexpress Co. Ltd. (Monmouth Junction, NJ, USA).

Techniques: Concentration Assay

Journal:

Article Title: Substantial depletion of the intracellular Ca 2+ stores is required for macroscopic activation of the Ca 2+ release-activated Ca 2+ current in rat basophilic leukaemia cells

doi: 10.1111/j.1469-7793.2000.t01-1-00247.x

Figure Lengend Snippet: A, the effect of dialysing individual cells with either 0.1 mM (low buffer) or 10 mM EGTA (high buffer) together with 30 μM IP3 and 2 mM ATP is shown. ICRAC amplitude (measured at −80 mV from voltage ramps from −100 to + 100 mV over 50 ms from a holding potential of 0 mV) is plotted against time after the onset of whole-cell recording. Whilst most of the cells dialysed with low buffer and IP3 (A1), IP3-F (A3), ionomycin (A4) or IP3 and ionomycin (A5) failed to generate any detectable ICRAC, the current was routinely activated in the presence of the SERCA pump blocker thapsigargin alone (A5). Dialysis with IP3-F and 10 mM EGTA activated ICRAC rapidly and to the maximal extent and the current was indistinguishable from that seen with IP3 and 10 mM EGTA (data not shown). IP3 interacted synergistically with the structurally distinct SERCA blockers thapsigargin (A2), thapsigargicin (A3) and cyclopiazonic acid (CPA) (A4) to generate ICRAC. The effect of SERCA pump blockers was dose dependent, as is shown in A2. All traces where macroscopic ICRAC is detected are fitted by a single exponential function. Insets show the I–V relationship of the current for each condition. The arrows in the plot of ICRAC development against time indicate where the inset I–V relationships were taken. B, ICRAC amplitude is plotted as a function of delay (B 1), time to peak (B2) and the time constant (τ) of the exponential fits (B3). The different conditions are stated in panel B 1. Each point represents mean ±s.e.m. of more than five cells. Delay was not significantly different between the conditions (P > 0.05). Time to peak and τ were significantly different depending whether (A1-A4) or not (A5) IP3 was present in the dialysing solution (P < 0.0001). In panel B 4, the rate of current development is plotted as a function of time for the conditions where ICRAC was activated by IP3 and thapsigargin (A2, n = 16) or thapsigargin alone (A5, n = 6). The first derivative of the exponential fit for each trace in either condition was calculated, multiplied by −1 and then pooled to calculate mean ±s.d. of the ICRAC rate.

Article Snippet: Depending on the experiment (described in the text), the Ca 2+ chelators EGTA (Sigma), BAPTA tetracaesium salt (BAPTA) or dimethyl BAPTA tetrapotassium salt (both from Molecular Probes) were added to this solution at the specified concentrations, as sometimes was 30 μm IP 3 , 2 μM thapsigargin (from three independent sources: Sigma, Calbiochem, Alomone Labs), 100 μM cyclopiazonic acid, or 2 μM thapsigargicin (Calbiochem).

Techniques: