Journal: bioRxiv

Article Title: Plakoglobin supports length-dependent force generation and cardiac resistance to fasting

doi: 10.64898/2026.10.09.757844

Figure Lengend Snippet: (A) Schematic representation of systemic AAV administration in mice. Six weeks after AAV injection animals were sacrificed and muscles were dissected. (B) Efficient AAV delivery into mouse skeletal muscle. Left: TA muscle sections from AAV injected mice were stained with antibodies against eGFP (bar, 200 μm). Right: RT-PCR of mRNA preparations from AAV-infected muscles using specific primers for plakoglobin ( JUP) . n = 4 mice per condition. Data are mean ± SEM. **, p<0.01 vs . shLacz by two-tailed unpaired Student t test. (C) Force–calcium curves of skinned skeletal muscle fibers from WT and plakoglobin knockdown muscle fibers at 2.1 µm sarcomere length. n = 3 mice per condition. (D) Force–calcium curves of skinned skeletal muscle fibers from WT and plakoglobin KD muscle fibers at 2.4 µm sarcomere length. n = 3 mice per condition. (E-F). Maximum calcium-activated tension (T max ; E) and calcium sensitivity (EC 50 ; F) of skeletal muscle fibers were analyzed. For WT, n=14 fibers at 2.1 μm and n=13 fibers at 2.4 μm; for plakoglobin knockdown (JUP-KD), n= 15 fibers at each sarcomere length. Fibers were obtained from n = 3 mice per group. Violin plots show individual fiber measurements; open circles represent the mean for each mouse, and horizontal lines and error bars represent mean ± SEM. WT and JUP-KD fibers were compared separately at each sarcomere length using two-sided nested t tests implemented as linear mixed-effects models, with genotype as a fixed effect and mouse as a random effect to account for multiple fibers measured within each mouse. Exact p values are shown.

Article Snippet: AAV vectors encoding shRNA targeting scrambled shLacz or the 3’UTR of mouse plakoglobin ( JUP ; gene ID 16480) under U6 promoter (experimentally validated shRNA sequence by manufacturer to knock down plakoglobin in cultured cells) were purchased from Vector BioLabs (Malvern, PA, USA) (Cat shAAV-262554) and were of Serotype 9, a serotype with a proven tropism for skeletal muscle cells ( ).

Techniques: Injection, Muscles, Staining, Reverse Transcription Polymerase Chain Reaction, Infection, Two Tailed Test, Knockdown, Sensitivity

Journal: bioRxiv

Article Title: Plakoglobin supports length-dependent force generation and cardiac resistance to fasting

doi: 10.64898/2026.10.09.757844

Figure Lengend Snippet: (A) Downregulation of plakoglobin reduces DGC stability and insulin signaling in skeletal muscle. Left: soluble fractions from AAV infected TA muscles were analyzed by immunoblotting. Right: densitometric measurements of the shown blots. n = 3 mice per condition. Data are mean ± SEM. *, p<0.05 and **, p<0.01 vs . shLacz by one-tailed unpaired Student t test. (B) Downregulation of plakoglobin in normal TA muscle activates MuRF1 expression, but not in the heart. RT-PCR of mRNA preparations from AAV-infected muscles using specific primers for plakoglobin ( JUP) and MuRF1. n = 4 mice per condition. Data are mean ± SEM. ns, not statistically significant. **, p<0.01 vs . shLacz by two-tailed unpaired Student t test. (C) Left: soluble fractions from AAV infected cardiac muscles were analyzed by immunoblotting. Right: densitometric measurements of the shown blots. n = 3 mice per condition. Data are mean ± SEM. ns, not statistically significant vs . shLacz by two-tailed unpaired Student t test.

Article Snippet: AAV vectors encoding shRNA targeting scrambled shLacz or the 3’UTR of mouse plakoglobin ( JUP ; gene ID 16480) under U6 promoter (experimentally validated shRNA sequence by manufacturer to knock down plakoglobin in cultured cells) were purchased from Vector BioLabs (Malvern, PA, USA) (Cat shAAV-262554) and were of Serotype 9, a serotype with a proven tropism for skeletal muscle cells ( ).

Techniques: Stability, Infection, Muscles, Western Blot, One-tailed Test, Expressing, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test

Journal: Autophagy Reports

Article Title: TRPML3 and TRPML1/3 differentially regulate unconventional MIF secretion through TMED10 and secretory autophagy

doi: 10.1080/27694127.2026.2740425

Figure Lengend Snippet: Relationship between MIF and TMED10. (A) Secretion of MIF was measured in HEK293T cells either untreated or treated with brefeldin A. (B) Secretion of MIF was measured in HEK293T cells untreated or treated with the indicated doses of probenecid. (C) GST or the indicated GST-TMED10 domains coupled to glutathione sepharose beads were incubated with whole cell lysates from HEK293T cells expressing Flag-MIF. Elutes were subjected to SDS-PAGE and immunoblotted with an anti-Flag antibody. (D) GST-MIF coupled to glutathione sepharose beads was incubated with whole cell lysate from HEK293T cells expressing GFP-TMED10 or GFP-TMED10(ΔCT). Elutes were subjected to SDS-PAGE and immunoblotted with anti-GFP antibody. (E) Secretion of MIF was measured in HEK293T cells expressing mock or GFP-TMED10. (F) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting TMED10 for 72 h. (G) A crosslink assay was performed in HEK293T cells expressing GFP-TMED10 with mock or Flag-MIF to examine the oligomeric form of TMED10. (H) Secretion of Flag-MIF was measured in HEK293T cells expressing mock, RAB1A, 1B, and 2A.

Article Snippet: For knockdown experiments, siRNA targeting human TMED10 (BIONEER, 10,972–1), human ATG5 (BIONEER, 9474–1), human SYT5 (BIONEER, 6861–1), and a negative control scrambled siRNA (Santa Cruz Biotechnology, sc-37007) were used.

Techniques: Incubation, Expressing, SDS Page, Control

Journal: Autophagy Reports

Article Title: TRPML3 and TRPML1/3 differentially regulate unconventional MIF secretion through TMED10 and secretory autophagy

doi: 10.1080/27694127.2026.2740425

Figure Lengend Snippet: Effect of autophagy on MIF secretion. (A) HeLa cells expressing GFP-LC3 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (B) Secretion of MIF was measured in HEK293T cells either untreated or treated with 10 μM SAR405. (C) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting ATG5 for 72 h. (D) Secretion of MIF was measured in HEK293T cells either untreated or treated with 100 nM bafilomycin A1. (E) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting SYT5 for 48 h. Knockdown efficiency was confirmed by immunoblotting for SYT5. (F) Secretion of Flag-MIF was measured in HEK293T cells expressing mock or GFP-SEC22B. (G) HeLa cells expressing GFP-SEC22B and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (H, upper) HeLa cells co-expressing GFP-MIF and Flag-VAMP4 were treated with cold digitonin (50 µg/ml), immunostained with an anti-Flag antibody, and imaged by fluorescence microscopy. (H, lower) HeLa cells expressing GFP-VAMP8 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy.

Article Snippet: For knockdown experiments, siRNA targeting human TMED10 (BIONEER, 10,972–1), human ATG5 (BIONEER, 9474–1), human SYT5 (BIONEER, 6861–1), and a negative control scrambled siRNA (Santa Cruz Biotechnology, sc-37007) were used.

Techniques: Expressing, Fluorescence, Microscopy, Control, Knockdown, Western Blot