Journal: JACC: Basic to Translational Science

Article Title: JP2 and JCN Crosstalk Abrogate MURF1-Mediated JCN Ubiquitination and Degradation in Cardiomyocytes

doi: 10.1016/j.jacbts.2026.101534

Figure Lengend Snippet: Ca 2+ Handling in Adult Mouse Cardiomyocytes (A to C) Adult mouse cardiomyocytes were infected with Ad-JP2 or Ad-HA and exposed to palmitate or oleate. RyR2 Ca 2+ leak (F/F0), caffeine-induced peak (F/F0), and Tau-SERCA were measured. (D to F) Adult mouse cardiomyocytes were infected with Ad-JCN or Ad-HA and exposed to palmitate or oleate. RyR2 Ca 2+ leak (F/F0), Caffeine-induced peak (F/F0), and Tau-SERCA were detected. (G to I) Adult mouse cardiomyocytes were infected with Ad-shJP2 or Ad-HA. RyR2 Ca 2+ leak (F/F0), caffeine-induced peak (F/F0), and Tau-SERCA were detected. (J to L) Adult mouse cardiomyocytes were infected with Ad-AsJCN or Ad-HA. RyR2 Ca 2+ leak (F/F0), caffeine-induced peak (F/F0), and Tau-SERCA were determined. Data are mean ± SD, n = 9-11 different adult mouse cardiomyocytes in each group. Two-way analysis of variance followed by Newman-Keuls post hoc test was performed for multiple pairwise comparisons. Student's t -test was used for comparison between 2 groups. ∗ P < 0.05 vs Ad-HA or oleate + Ad-HA, and † P < 0.05 vs palmitate + Ad-HA. ∗ and †: P < 0.05; ∗∗ and ††: P < 0.01; ∗∗∗ and †††: P < 0.001. Abbreviations as in .

Article Snippet: Then, sarcoplasmic reticulum (SR) Ca 2+ leak via RyRs was estimated in 0 Ca 2+ /0 Na + Tyrode's solution containing tetracaine (1 mmol/L, MedChemExpress, HY-A0079).

Techniques: Infection, Comparison

Journal: JACC: Basic to Translational Science

Article Title: JP2 and JCN Crosstalk Abrogate MURF1-Mediated JCN Ubiquitination and Degradation in Cardiomyocytes

doi: 10.1016/j.jacbts.2026.101534

Figure Lengend Snippet: Schematic Mechanism In normal conditions, JP2 binds to JCN and blocks the MURF1-JCN interaction, thereby preventing MURF1-mediated ubiquitination and proteasome-dependent degradation of JCN. However, the protein level of JP2 is reduced under stress, enabling MURF1 to bind to JCN and promoting MURF1-mediated ubiquitination and degradation of JCN. JCN reduction triggers the excessive opening and Ca 2+ release from RyR2, leading to cardiomyocyte injury. In contrast, overexpression of JP2 or JCN protects intracellular Ca 2+ homeostasis and prevents cell death under stress.

Article Snippet: Then, sarcoplasmic reticulum (SR) Ca 2+ leak via RyRs was estimated in 0 Ca 2+ /0 Na + Tyrode's solution containing tetracaine (1 mmol/L, MedChemExpress, HY-A0079).

Techniques: Ubiquitin Proteomics, Over Expression

Journal: STAR Protocols

Article Title: Protocol for differentiation and efficient AAV-mediated gene delivery to hiPSC-derived microglia for functional studies

doi: 10.1016/j.xpro.2026.104455

Figure Lengend Snippet: Propagation of calcium signal within microglia after ATP stimulation (A) Baseline GCaMP8s expression. (B) Regions of interest (ROIs): one somatic (ROI 1) and two distal regions (ROI 2 and 3) were chosen. (C) Snapshots showing propagation of the fluorescence signal within the cell following ATP stimulation. (D) Normalized fluorescence traces (ΔF/F0, F0 = mean fluorescence intensity over 10 s prior to stimuli) recorded from ROIs in B. The period shaded in green indicates when ATP was present in the recording chamber. ROI 3 (most distal) exhibits spontaneous activity prior to stimulation, indicated by asterisks. Dashed vertical lines indicate time points (t0-t3) corresponding to images in C.

Article Snippet: Dulbecco’s phosphate buffered saline without Ca 2+ and Mg 2+ , DPBS (−/−) , Thermo Fisher Scientific , 14190–086.

Techniques: Expressing, Fluorescence, Activity Assay

Journal: STAR Protocols

Article Title: Protocol for differentiation and efficient AAV-mediated gene delivery to hiPSC-derived microglia for functional studies

doi: 10.1016/j.xpro.2026.104455

Figure Lengend Snippet:

Article Snippet: Dulbecco’s phosphate buffered saline without Ca 2+ and Mg 2+ , DPBS (−/−) , Thermo Fisher Scientific , 14190–086.

Techniques: Virus, Recombinant, Saline, Plasmid Preparation, Expressing, Software, Hood, Sterility, Electron Microscopy, Inverted Microscopy, Flow Cytometry, Microscopy, Cell Culture, Fluorescence, Imaging, Dispersion

Journal: Frontiers in Pharmacology

Article Title: The TASK-1 and TASK-3 activator JG-C3-98 attenuates cold and mechanical responses in primary somatosensory neurons

doi: 10.3389/fphar.2026.1844406

Figure Lengend Snippet: JG-C3-98 shifts the cold threshold to lower temperatures and reduces the mechanical responses of primary sensory neurons in culture. (A) Representative traces of [Ca 2+ ] i (as F 340 /F 380nm ratio) in cold-sensitive neurons with and without JG-C3-98 treatment. Two cold pulses were applied in control solution (left panel), while in the treatment condition, 100 µM JG-C3-98 was applied prior to and during the 2 nd pulse (right panel). Arrowheads indicate the thermal thresholds and maximal responses (control black, JG-C3-98 red). (B,C) Dot plots summarizing the individual and mean cold threshold (±SEM) and the individual and mean maximal response (±SEM) of cold-sensitive neurons in control condition (n = 27, n.s. p = 0.7027 and n.s. p = 0.0642, respectively), or treated with 100 µM JG-C3-98 (n = 33, ***p = 0.00003 and *p = 0.0133, respectively). (D) Representative traces of [Ca 2+ ] i (as F 340 /F 380nm ratio) in mechano-sensitive neurons with and without JG-C3-98. Two pulses of 210 mOsm/L (hypoosmotic) solution were applied in control condition (left panel) and in the presence of 100 µM JG-C3-98 before and during the second pulse (right panel). Arrowheads indicate the maximal responses (control black, JG-C3-98 red). (E,F) Dot plots showing the individual and mean maximal response (±SEM) to hypoosmotic solution in cultured neurons, sensitive and insensitive to 100 µM AITC (n = 13, n.s. p = 0.8262 in control, and n = 10, *p = 0.0212 in JG-C3-98, for AITC-sensitive neurons; and n = 12, n.s. p = 0.4522 in control, and n = 26, ***p = 0.0009 in JG-C3-98, for AITC-insensitive neurons). Filled and open blue triangles indicate the mean amplitude of the AITC-evoked response in AITC-sensitive and AITC-insensitive neurons, respectively. Statistical analysis was carried out using two tailed Student’s t-test for paired samples (n.s., p > 0.05; *p < 0.05; ***p < 0.001).

Article Snippet: For ratiometric Ca 2+ imaging experiments, sensory neurons were incubated in 5 μM Fura-2 AM (F1221, Invitrogen-Thermo Fisher Scientific, Waltham, MA, United States of America) in standard extracellular solution (containing in mM: 140 NaCl, 3 KCl, 1.3 MgCl 2 , 2.4 CaCl 2 , 10 HEPES, 10 glucose; 298 mOsm/kg, pH 7.4 adjusted with NaOH), supplemented with 0.02% Pluronic acid (P6867, Invitrogen-Thermo Fisher Scientific, Waltham, MA, United States of America) for 50 min at 37 °C, in darkness.

Techniques: Control, Cell Culture, Two Tailed Test