Journal: bioRxiv

Article Title: Phosphorylation of the rod-tail hinge region of cingulin regulates its interaction with nonmuscle myosin-2B

doi: 10.64898/2026.04.02.716052

Figure Lengend Snippet: Inhibition of either CK1 or CK2 promotes the rescue of junctional NM2B in cingulin-KO cells expressing the depho-6 mutant of cingulin. (A) Top: scheme of GFP-tagged canine CGN (cCGN-FL), with the GFP tag (green), globular head (gray), coiled-coil rod (white) and globular tail (blue) domains (amino-acid residue boundaries are indicated above the scheme). Bottom: C-terminal sequences of WT canine CGN (cCGN-FL, sequence 1140–1190 with specific residues indicated in the region) and corresponding sequences of the cCGN-dephosphomimetic-6 mutant (depho-6), which does not rescue junctional NM2B (Fig. S1K’). (B-G) IF microscopy analysis and localization (left) and quantification of junctional labeling (relative fluorescence intensity) (right) of NM2B in CGN-KO MDCK cells rescued with GFP-cCGN-FL (B, D and F), or with GFP-cCGN-depho-6 mutant (C, E and G) treated either with DMSO (B and C) or with CK1 inhibitor (D and E: 25 μM, 8h) or with CK2 inhibitor (F and G: 25 μM, 14h). Arrows and arrowheads show increased, normal and decreased/undetected junctional labeling, respectively. Quantifications of relative fluorescent intensity (RFI) shows the ratio between the junctional staining of NM2B versus the junctional marker PLEKHA6 (n=70 junctions) from three independent experiments. Data in quantifications are represented as mean±SD. Statistical significance was determined by unpaired Mann-Whitney’s test. ***p≤0.001. Scale bar (G)= 10 μm.

Article Snippet: Drugs treatments were as follows (final concentration, duration, catalog number and source): CK1 inhibitor (25 μM, 8 h, D4476 MedChemExpress) and CK2 inhibitor (5 μM, 14 h, CX4945 Selleckchem).

Techniques: Inhibition, Expressing, Mutagenesis, Residue, Sequencing, Microscopy, Labeling, Fluorescence, Staining, Marker

Journal: iScience

Article Title: CK1δ-dependent SNAPIN dysregulation drives lysosomal failure in HIV-1 Vpr-exposed neurons: A targetable mechanism in HAND

doi: 10.1016/j.isci.2025.114544

Figure Lengend Snippet: HIV-1 Vpr increases neuronal CK1δ expression Immunofluorescence of CK1δ (green) in SH-SY5Y-derived neurons treated with vehicle, Vpr, LH846, or Vpr + LH846. LAMP1 (red) labels lysosomes; nuclei are DAPI-stained (blue). Representative qualitative images are shown. Scale bars: 10 µm.

Article Snippet: Anti-CK1δ antibody , Cell Signaling Technology , #2655; RRID: AB_2283593.

Techniques: Expressing, Immunofluorescence, Derivative Assay, Staining

Journal: iScience

Article Title: CK1δ-dependent SNAPIN dysregulation drives lysosomal failure in HIV-1 Vpr-exposed neurons: A targetable mechanism in HAND

doi: 10.1016/j.isci.2025.114544

Figure Lengend Snippet: CK1δ inhibition restores lysosomal distribution disrupted by HIV-1 Vpr SH-SY5Y neurons expressing LAMP1-mCherry and SNAPIN wild-type (WT) or S50D were treated with Vpr ± LH846 (1.5 μM). Radial intensity profiles quantified lysosomal distance from nuclei. Scale bars: 10 μm.

Article Snippet: Anti-CK1δ antibody , Cell Signaling Technology , #2655; RRID: AB_2283593.

Techniques: Inhibition, Expressing

Journal: iScience

Article Title: CK1δ-dependent SNAPIN dysregulation drives lysosomal failure in HIV-1 Vpr-exposed neurons: A targetable mechanism in HAND

doi: 10.1016/j.isci.2025.114544

Figure Lengend Snippet: CK1δ inhibition restores SNAPIN distribution and lysosomal positioning (A) Representative immunofluorescence images of neurons expressing SNAPIN-GFP and LAMP1-mCherry treated with Vpr ± LH846. Images show SNAPIN relocalization following CK1δ inhibition. (B) Quantification of SNAPIN–LAMP1 colocalization using Manders’ coefficient. Colocalization increased with Vpr and was normalized by LH846 (mean ± SEM; n ≥ 100 cells from ≥3 independent differentiations). Statistics: one-way ANOVA with Tukey post hoc; p < 0.05. Scale bars: 10 µm.

Article Snippet: Anti-CK1δ antibody , Cell Signaling Technology , #2655; RRID: AB_2283593.

Techniques: Inhibition, Immunofluorescence, Expressing

Journal: iScience

Article Title: CK1δ-dependent SNAPIN dysregulation drives lysosomal failure in HIV-1 Vpr-exposed neurons: A targetable mechanism in HAND

doi: 10.1016/j.isci.2025.114544

Figure Lengend Snippet: CK1δ inhibition restores lysosomal acidification and mitophagy in Vpr-exposed neurons (A) Lysosomal pH quantified using pHluorin-mCherry-LC3 (mean ± SEM; n ≥ 100 cells/condition from ≥3 independent differentiations). One-way ANOVA: F(3, 8.8) = 58.2, p < 0.001. (B) Representative confocal images of LAMP1 (red) and MitoGreen (green). Scale bars: 10 μm. (C) Colocalization (Manders tM2) between LAMP1 and MitoGreen (mean ± SEM; n ≥ 100 cells/condition from ≥3 differentiations). Welch ANOVA: F(3, 8.1) = 9.15, p = 0.006; Tukey post hoc p < 0.001 for Vpr vs. control and Vpr vs. Vpr + LH846.

Article Snippet: Anti-CK1δ antibody , Cell Signaling Technology , #2655; RRID: AB_2283593.

Techniques: Inhibition, Control