Journal: eLife

Article Title: An altered cell-specific subcellular distribution of translesion synthesis DNA polymerase kappa (POLK) in aging mouse neurons

doi: 10.7554/eLife.101533

Figure Lengend Snippet: ( A ) Schematic of cytoplasmic compartments and condensates and respective markers in parentheses that were assayed to colocalize with cytoplasmic POLK. ( B ) Cytoplasmic POLK (green) expression colocalizing with G3BP1 and LAMP1 (red) in fluorescent-nissl stained cells (blue) of mouse brain tissue in 18-month-old brain. Line scan in the boxed region shows POLK colocalizing with LAMP1 and G3BP1. ( C – F ) Cytoplasmic POLK (green), EEA1, CTSB, CTSD, and GBA1 (red) in fluorescent-nissl stained cells (blue) of mouse brain tissue in 18-month-old brain. Line scan in the boxed region shows level of POLK colocalization with the proteins. Highest colocalization with CTSD, partial with EEA1 and CTSB, and minimal GBA1.

Article Snippet: Antibody , Rat monoclonal anti-1D4B (LAMP1) , DSHB , RRID: AB_2134500 , IF (1:200).

Techniques: Expressing, Staining

Journal: eLife

Article Title: An altered cell-specific subcellular distribution of translesion synthesis DNA polymerase kappa (POLK) in aging mouse neurons

doi: 10.7554/eLife.101533

Figure Lengend Snippet: ( A ) Cytoplasmic POLK (green) expression colocalizing with G3BP1 (blue) in fluorescent-nissl stained cells (purple) of mouse brain tissue from M1 and S1 cortical regions in 18-month-old brain but not in young 1 month. ( B ) Cytoplasmic POLK (green) expression colocalizing with LAMP1 (blue) in fluorescent-nissl stained cells (purple) of mouse brain tissue from M1 and S1 cortical regions of 18-month brain. ( C ) Additional representative image with channel separation for immunofluorescence staining of wild-type mouse brain cortical areas S1, showing cytoplasmic POLK is colocalized with stress granule marker G3BP1 and endo/lysosomal marker LAMP1. Arrows indicate few representative sites of colocalization in both images.

Article Snippet: Antibody , Rat monoclonal anti-1D4B (LAMP1) , DSHB , RRID: AB_2134500 , IF (1:200).

Techniques: Expressing, Staining, Immunofluorescence, Marker

Journal: Bioactive Materials

Article Title: Countering postoperative immune suppression with a self-assembling dendritic cell nanovaccine

doi: 10.1016/j.bioactmat.2026.05.005

Figure Lengend Snippet: BAITs promote DC activation and antigen presentation via enhanced lysosomal processing. (A) Schematic for MHC Ⅰ/Ⅱ immunostaining. DCs were treated with antigens or BAITs (12 h), then incubated in fresh medium (with TGFβ) for 24 h before staining. (B, C) Quantification of surface MHC Ⅰ (B) and MHC Ⅱ (C) on DCs (n = 8; n.s. is P > 0.05, ∗∗∗ is P < 0.001, ∗∗∗∗ is P < 0.0001 by one-way ANOVA with Bonferroni post-hoc test). (D, E) Representative immunofluorescence images of DCs (blue: nuclei; yellow: LAMP1; green: MHC Ⅰ; magenta: MHC Ⅱ). BAIT-treated DCs show strong surface MHC and minimal intracellular antigen signal, whereas antigen-treated DCs show retained antigen (red) and weak surface MHC signal. (F–H) Flow cytometric analysis of MHC Ⅰ (F), MHC Ⅱ (G), and CD80 (H) expression on murine CD11c + DCs after exposure to antigens or BAITs. (I) Quantification of MHC Ⅰ, MHC Ⅱ, and CD80 expression in DCs by flow cytometry (n = 3; ∗∗∗ is P < 0.001, ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (J) Heatmap of DC activation-related gene expression after treatment with antigens or BAITs, highlighting upregulation of maturation and antigen presentation genes and downregulation of immunosuppressive genes by BAITs (n = 3). Data are presented as mean ± SD.

Article Snippet: After treatment, the cells were washed twice with PBS and fixed in 4% paraformaldehyde for 20 min. For intracellular protein staining, fixed cells were permeabilized with 0.1% Triton X-100 at room temperature for 10 min. Next, the cells were blocked with 2% BSA and incubated with rat CoraLite Plus 488 anti-mouse LAMP1 antibody (1:200) and rabbit anti-mouse pSAP (1:200) antibody overnight at 4 °C.

Techniques: Activation Assay, Immunopeptidomics, Immunostaining, Incubation, Staining, Immunofluorescence, Expressing, Flow Cytometry, Gene Expression

Journal: The EMBO Journal

Article Title: TRPML1 suppresses pulmonary fibrosis by limiting collagen and elastin deposition

doi: 10.1038/s44318-026-00712-4

Figure Lengend Snippet: ( A ) Fura-2 Ca 2+ imaging experiments using HEK293 cells expressing plasma membrane (PM) variants of human TRPML1 and TPC2, or TRPML3, respectively, indicating the specific levels of activation. Channels were stimulated with either ML-SA1, MK6-83, or a range of TRPML1 agonists obtained from CasmaTherapeutics, in the concentration of 10 µM, each with the exception of ML1-SA1, which was applied at a concentration of 30 µM, ( B ) Average values (mean ± SEM) from 7 to 10 independent experiments per condition are shown with up to ten cells, each. Statistical significance was assessed using two-way ANOVA followed by Tukey’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Exact p values were: Comparison of TRPML1 treated with ML-SA1 10 µM to WR1-002 10 µM, p = 0.0130; TRPML1 treated with MK6-83 10 µM to WR250-003 10 µM, p = 0.0314; TRPML1 treated with MK6-83 10 µM to WR1-002 10 µM, p = 0.0048; TRPML1 treated with ML1-SA1 30 µM to WR250-003 10 µM, p = 0.0003; TRPML1 treated with ML1-SA1 30 µM to WR1-002 10 µM, p < 0.0001; TRPML1 treated with WR250-003 10 µM to TRPML3 treated with WR250-003 10 µM, p < 0.0001; TRPML1 treated with WR1-002 10 µM to TRPML3 treated with WR1-002 10 µM, p < 0.0001. ( C ) Cartoon illustrating the LAMP1 translocation assay as used in ( D – H ). Upon lysosomal exocytosis, the lysosomal protein LAMP1 is detected on the plasma membrane (PM) by anti-LAMP1 antibody, followed by visualization with Alexa Fluor 488-conjugated secondary antibody. Purple dots shall represent lysosomal luminal content (e.g., MMPs) being released. ( D – G ) Lysosomal exocytosis experiments measuring LAMP1 translocation in WT and Trpml1 −/− alveolar macrophages (AMΦ). Maximum effects were obtained with ionomycin (4 µM). Shown are results (mean ± SEM) obtained after 120 min treatment with DMSO, ML-SA1 as control and additional compounds as indicated. Treatment time with ionomycin (4 µM) was 10 min. Representative images of LAMP1 translocation using WT and Trpml1 −/− AMΦ are shown in ( D , G ). Shown in ( E , F ) are normalized mean values ± SEM from at least four independent experiments, each. Statistical significance was assessed using two-way ANOVA followed by Dunnett’s post hoc test. **** p < 0.0001. For all listed compounds in ( F ), WT samples showed a statistically significant difference compared to the DMSO control, with an exact p < 0.0001. In KO samples, all compounds were not significant except for the positive control ( p < 0.0001). For the compounds listed in panel ( E ), most showed statistically significant differences compared to DMSO control in WT samples ( p < 0.0001), except for WR1-001 5 µM and WR250-003 0.7 µM, which were not significant. In KO samples, all compounds were not significant except for the positive control, which was significant ( p < 0.0001). .

Article Snippet: LAMP1 (rat) , Santa Cruz , sc-19992.

Techniques: Imaging, Expressing, Clinical Proteomics, Membrane, Activation Assay, Concentration Assay, Comparison, Translocation Assay, Control, Positive Control