Journal: bioRxiv

Article Title: Regulation of spontaneous neurotransmission and homeostatic synaptic plasticity by synaptotagmin-1 disease variants at the SNARE primary interface

doi: 10.64898/2026.02.17.706274

Figure Lengend Snippet: (A) Representative raw traces of mEPSCs of Empty Vector (CTRL) and Syt1 variant (N341S) neurons following acute 1 hr treatment with vehicle (VEH), TTX, or TTX+AP5 treatment. (B) Comparison of mean baseline mEPSC amplitude (pA) for each group after incubations (Two-way ANOVA with Dunnett’s multiple comparisons test: VEH vs. TTX of CTRL p=0.120; VEH vs. TTX+APV of CTRL p=0.006; VEH vs. TTX of N341S p=0.050; VEH vs. TTX+APV of N341S p=0.796, N=2, n=6-7 per group). (C) Comparison of mean baseline mEPSC frequency (Hz) for each group after incubations (Two-way ANOVA with Dunnett’s multiple comparisons test: VEH vs. TTX of CTRL p=0.986; VEH vs. TTX+APV of CTRL p=0.991; VEH vs. TTX of N341S p=0.519; VEH vs. TTX+APV of N341S p=0.276, N=2, n=6-7 per group). (D) Representative raw traces of mEPSCs of empty vector (CTRL) and Syt1 variant (N341S) neurons after chronic 48 hr DMSO and cAMPS-RP treatment. (E) Comparison of mean baseline mEPSC amplitude (pA) for each group after incubations (Two-way ANOVA with Sidak’s multiple comparisons test: DMSO vs. CAMPS-RP of CTRL p=0.007; DMSO vs. cAMPS-RP of N341S p=0.865; N=2-3, n=7-14 per group). (F) Comparison of mean baseline mEPSC frequency (Hz) for each group after incubations (Two-way ANOVA with Sidak’s multiple comparisons test: DMSO vs. CAMPS-RP of CTRL p=0.170; DMSO vs. cAMPS-RP of N341S p=0.856; N=2-3, n=7-14 per group).

Article Snippet: To model homeostatic plasticity using chronic pharmacological effect, neurons were incubated for 48 hours with cAMPS-RP (MedChemExpress).

Techniques: Plasmid Preparation, Variant Assay, Comparison

Journal: bioRxiv

Article Title: Diet-induced macrophage-driven inflammation fuels pancreatic plasticity and aggressiveness

doi: 10.64898/2026.02.10.703309

Figure Lengend Snippet: A) Quantification of P2rx7 expression by RT-qPCR in control (Ctrl) murine cell lines treated with fatty acids (FAs) (linolenic acid, LA [200µM]; oleic acid, OA [50µM]; palmitic acid, PA [50µM sodium palmitate]). Shown is the mean ± SEM (n=4 cell lines in duplicates for LA, and 3 cell lines in triplicate for OA and PA, P values as determined by unpaired T-test). B) Quantification of P2RX7 expression by RT-qPCR in PDX-derived primary human cell lines treated with FAs. Shown is the mean fold change ± SEM with controls set as 1.0 (n=3 cell lines, P values as determined by unpaired T-test). C) Quantification of P2rx7 expression by RT-qPCR in Ctrl murine cell lines treated with adipose tissue conditioned medium (ACM) for 48 hours. Shown is the mean ± SEM (n=4 cell lines in duplicates, P values as determined by unpaired T-test). D) Dot plot showing the correlation between P2RX7 and CAMP expression in TCGA patients. P values and r were calculated by Pearson’s correlation. E) Quantification of Camp expression by RT-qPCR in immortalized bone marrow-derived macrophages treated with FAs (linolenic acid, LA; oleic acid, OA; palmitic acid, PA). Shown is the mean ± STDEV (n=3, P values as determined by unpaired T-test). F) Quantification of CAMP positive total cells (left panel) and CAMP positive cells in macrophages (F4/80) (right panel) from the bone marrow determined by flow cytometry in mice fed with Ctrl diet and HFD. Shown is the mean ± STDEV (n=3 mice for control group, 4 for HFD group, P values as determined by unpaired T-test). G) Quantification of CAMP positive tumor-associated macrophages determined by flow cytometry in mice fed with Ctrl diet and HFD. Shown is the mean ± STDEV (n=3 mice, P values as determined by unpaired T-test). H) Quantification of P2rx7 expression by RT-qPCR in Ctrl murine cell lines treated with macrophage conditioned medium (MCM) or fatty acid-trained macrophage conditioned medium (F-MCM) for 48 hours. Shown is the mean fold change ± SEM with Ctrl set as 1.0 (n=3 cell lines in triplicates, P values as determined by One Way ANOVA). I) Quantification of Epcam expression by RT-qPCR in Ctrl murine cell lines treated with MCM or F-MCM for 48 hours. Shown is the mean fold change ± SEM with Ctrl set as 1.0 (n=3 cell lines in duplicates, P values as determined by One Way ANOVA). J) Representative brightfield microscopy images of Ctrl cells treated with MCM and F-MCM. Scale bar= 50µm. K) Quantification of P2RX7 positive cells determined by flow cytometry in a Ctrl cell culture treated with recombinant (r) CAMP (100 µg/mL) for 48 hours. Shown is the mean ± SEM (n=8, P values as determined by unpaired T-test). L) Quantification of CXCR4 positive cells determined by flow cytometry in a Ctrl cell culture treated with rCAMP (100 µg/mL) for 48 hours. Shown is the mean ± SEM (n=6, P values as determined by unpaired T-test). M) Quantification by ELISA of soluble LL-37 levels in sera from healthy donors (Healthy) (n=17) and PDAC patients with resectable (R) (n=21), locally advanced (LA) (n=30) or metastatic disease (M) (n=37). Shown is the mean of log normalized concentration ± SEM (P values as determined by One-Way ANOVA, Dunnet’s test). N) Quantification by ELISA of soluble LL-37 levels in sera from PDAC patients from the highest and lowest quartiles of BMI (n=22 patients per condition). Shown is the mean of log normalized concentration ± SEM (P values as determined by unpaired T-test). O) Violin plot showing the quantification of CAMP expression in PDAC patients based on their BMI 1 year before diagnosis (n for <25=75, n for ≥25=89). P value as determined by Wilcoxon test. P) Violin plot showing the quantification of CAMP expression in PDAC patients based on their BMI at time of diagnosis (n for <25=35, n for ≥25=54). P value as determined by Wilcoxon test. Q) Violin plot showing the quantification of CAMP expression in PDAC patients based on their BMI transition (patients previously obese that lost their weight versus those that always present a BMI<25) (n for always <25=75, n for obese → lean =35). P value as determined by Wilcoxon test. P values: *, <0.05; **, <0.01; ***, <0.001; ****, <0.0001.

Article Snippet: For stimulation assays, tumor or immune cells were seeded in 6-well plates and treated with 1 ml of complete RPMI medium plus recombinant CAMP (5 μg/mL, Cat. no. HY-P4855, MedChemExpress) during 24h/48h, fatty acids (50 μM, oleic acid; 50 μM sodium palmitate; 200 μM linoleic acid; Cat no. O3008, P9767, L9530) or MØ-derived conditioned media.

Techniques: Expressing, Quantitative RT-PCR, Control, Derivative Assay, Flow Cytometry, Microscopy, Cell Culture, Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay, Biomarker Discovery

Journal: bioRxiv

Article Title: Diet-induced macrophage-driven inflammation fuels pancreatic plasticity and aggressiveness

doi: 10.64898/2026.02.10.703309

Figure Lengend Snippet: A) Quantification of Pglyrp1 expression in control (Ctrl) and high-fat diet (HFD) primary murine cell lines included in the RNA-seq analyses. Shown is the mean ± SEM (n=6, P values as determined by T-test). B) Quantification of PGLYRP1 positive cells treated with recombinant (r) CAMP (100 µg/mL) for 48 hours. Shown is the mean ± SEM (n=6, P values as determined by unpaired T-test). C) Representative flow cytometry plots showing YFP positive control (Ctrl) and high-fat diet (HFD) primary tumor cells co-cultured with primary bone marrow-derived macrophages stained with F4/80. Live double positive events count as phagocytosed cells. D) Quantification of the percentage of phagocytosed tumor cells by flow cytometry. Shown is the mean ± SEM (n=3 cell lines in triplicates, P values as determined by unpaired T-test). P values: *, <0.05; **, <0.01; ***, <0.001; ****, <0.0001.

Article Snippet: For stimulation assays, tumor or immune cells were seeded in 6-well plates and treated with 1 ml of complete RPMI medium plus recombinant CAMP (5 μg/mL, Cat. no. HY-P4855, MedChemExpress) during 24h/48h, fatty acids (50 μM, oleic acid; 50 μM sodium palmitate; 200 μM linoleic acid; Cat no. O3008, P9767, L9530) or MØ-derived conditioned media.

Techniques: Expressing, Control, RNA Sequencing, Recombinant, Flow Cytometry, Positive Control, Cell Culture, Derivative Assay, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: Overexpression of GPER1 suppressed esophageal carcinoma growth via activating cAMP pathway

doi: 10.4081/ejh.2026.4422

Figure Lengend Snippet: Inhibition of the cAMP/PKA pathway attenuates GPER1-induced ferroptosis in esophageal cancer cells. KYSE70 and KYSE150 cells were treated with or without the cAMP/PKA inhibitor H89 (10 μM) for 24 h under four conditions: negative control with vehicle (NC + Veh), negative control with H89 (NC + H89), GPER1 overexpression with vehicle (GPER1-OE + Veh), and GPER1 overexpression with H89 (GPER1-OE + H89). a,d ) Cell viability was determined by CCK-8 assay in KYSE70 ( a ) and KYSE150 ( d ) cells. b,e ) Intracellular ROS levels were measured by flow cytometry using a DCFH-DA probe in KYSE70 ( b ) and KYSE150 ( e ) cells. c,f ) Intracellular Fe²⁺ content was quantified by an iron assay kit in KYSE70 ( c ) and KYSE150 ( f ) cells. g ) Protein expression levels of the ferroptosis markers ACSL4 and GPX4, and the cAMP pathway components p-CREB1 and CREB1, were analyzed by Western blotting in both KYSE70 and KYSE150 cells; left panels show representative Western blot images, and the right panels show the corresponding quantitative densitometric analysis of the protein bands, normalized to GAPDH (for ACSL4 and GPX4) or total CREB1 (for p-CREB1). All quantitative data are presented as the mean ±SD from three independent experiments (n=3). Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test; * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001; ns, not significant.

Article Snippet: To functionally validate the role of the cAMP pathway in GPER1-induced ferroptosis, KYSE70 and KYSE150 cells with or without GPER1 overexpression were treated with the cAMP/PKA pathway inhibitor H89 (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Inhibition, Negative Control, Over Expression, CCK-8 Assay, Flow Cytometry, Iron Assay, Expressing, Western Blot