Journal: International Journal of Molecular Sciences

Article Title: Photoreceptor Compartment-Specific TULP1 Interactomes

doi: 10.3390/ijms22158066

Figure Lengend Snippet: IP of retinal lysate. The top panels show Western blot analysis of the MAP1B IP experimental samples probed with Tulp1 antibodies. In the IP product lane, a band corresponding to Tulp1 is detected. A corresponding band is seen in the rat retinal lysate and wt mouse retinal homogenate but not in the tulp1−/− retinal lysate, liver lysate or non-specific IgG IP lanes. The bottom panels show Western blot analysis of the MAP1B IP experimental samples probed with MAP1B antibodies. In the IP product, rat retinal lysate, wt mouse retinal lysate and tulp1−/− retinal lysate lanes, a band corresponding to MAP1B is detected. No bands are seen in the liver or non-specific IgG IP sample lanes.

Article Snippet: Slides were then incubated with blocking solution (1% BSA and 10% Donkey serum in freshly prepared 1X PBS) for 2 h at RT and then subsequently incubated with primary antibodies: MAP1B at 1:100 (N-19, Santa Cruz Biotechnology), Kif3a at 1:300 (Abcam, ab11259), Ribeye at 1:1000 (BD Transduction, 612044) and Tulp1 at 1:500 (mTulp1) [ ] (diluted in 1× PBS with 1% BSA) overnight at 4 °C.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Photoreceptor Compartment-Specific TULP1 Interactomes

doi: 10.3390/ijms22158066

Figure Lengend Snippet: Immunolocalization of MAP1B and Tulp1 in P17 mouse retinas. ( A ) Wt retinal sections stained with MAP1B (red) and Tulp1 (green). ( B ) Tulp1−/− retinal sections stained with MAP1B (red) and Tulp1 (green). Sections were counterstained with DAPI (blue). Scale bar: 50 µm. INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS, inner segment layer; OS, outer segment layer.

Article Snippet: Slides were then incubated with blocking solution (1% BSA and 10% Donkey serum in freshly prepared 1X PBS) for 2 h at RT and then subsequently incubated with primary antibodies: MAP1B at 1:100 (N-19, Santa Cruz Biotechnology), Kif3a at 1:300 (Abcam, ab11259), Ribeye at 1:1000 (BD Transduction, 612044) and Tulp1 at 1:500 (mTulp1) [ ] (diluted in 1× PBS with 1% BSA) overnight at 4 °C.

Techniques: Staining

Journal: Frontiers in Oncology

Article Title: CDK1-driven phosphorylation networks promote glioblastoma progression via MAP1B-mediated microtubule destabilization

doi: 10.3389/fonc.2025.1646698

Figure Lengend Snippet: CDK1-mediated MAP1B phosphorylation is positively correlated with the prognosis of GBM. (A) Overlap analysis of CPTAC phosphoproteome data (blue) with CDK1-regulated phosphosites (red). (B) Heatmap of MAP1B phosphosites in the shCDK1 and shCtrl groups. Red indicates upregulated values relative to the mean, while blue indicates downregulated values relative to the mean. The intensity of the color corresponds to the magnitude of the deviation. (C) Prognostic significance of MAP1B phosphorylation sites. Kaplan-Meier survival analysis of GBM patients stratified by phosphorylation levels at S832, S1260, S1899, S1939, S2209, S2271(log-rank test, p < 0.05). High phosphorylation correlates with poor overall survival (n=99). (D, E) CoIP showing interactions between CDK1 and MAP1B. (F) Detecting pS/T phosphorylation of MAP1B in shCDK1 and shCtrl U251 cells. (G) Detecting pS/T phosphorylation of MAP1B in overexpressed-CDK1(OE-CDK1) and empty vector(Vector) U251 cells.

Article Snippet: Phospho-(Ser/Thr) Phe Antibody (#9631, Cell Signaling Technology), MAP1B Polyclonal antibody(21633-1-AP, proteintech) and β-actin Polyclonal antibody(20536-1-AP, proteintech) were used for western blot.

Techniques: Phospho-proteomics, Plasmid Preparation

Journal: Frontiers in Oncology

Article Title: CDK1-driven phosphorylation networks promote glioblastoma progression via MAP1B-mediated microtubule destabilization

doi: 10.3389/fonc.2025.1646698

Figure Lengend Snippet: CDK1-mediated phosphorylation of MAP1B regulates microtubule stability in U251 cells. (A) Wound healing assays showing delayed gap closure in shMAP1B cells compared to shCtrl cells over 36 hours, ***p < 0.001 (t-test), ****p < 0.0001 (t-test). (B) Transwell migration assays confirmed the impaired migration ability of shMAP1B cells. ***p < 0.001 (t-test). (C) CCK-8 proliferation assays revealed a significant decrease in cell viability in shMAP1B cells ***p < 0.001 (t-test). (D) Wiki-pathways analysis of differently expressed phosphoproteins. (E) Representative photographs of alpha tubulin staining of in U251 cells. Scale bar: 20 μm. Cells were stained with an antibody against acetyl-α-tubulin (green). Nuclei were counterstained with DAPI (blue). Scale bar, 20 μm.

Article Snippet: Phospho-(Ser/Thr) Phe Antibody (#9631, Cell Signaling Technology), MAP1B Polyclonal antibody(21633-1-AP, proteintech) and β-actin Polyclonal antibody(20536-1-AP, proteintech) were used for western blot.

Techniques: Phospho-proteomics, Migration, CCK-8 Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: Photoreceptor Compartment-Specific TULP1 Interactomes

doi: 10.3390/ijms22158066

Figure Lengend Snippet: IP of retinal lysate. The top panels show Western blot analysis of the MAP1B IP experimental samples probed with Tulp1 antibodies. In the IP product lane, a band corresponding to Tulp1 is detected. A corresponding band is seen in the rat retinal lysate and wt mouse retinal homogenate but not in the tulp1−/− retinal lysate, liver lysate or non-specific IgG IP lanes. The bottom panels show Western blot analysis of the MAP1B IP experimental samples probed with MAP1B antibodies. In the IP product, rat retinal lysate, wt mouse retinal lysate and tulp1−/− retinal lysate lanes, a band corresponding to MAP1B is detected. No bands are seen in the liver or non-specific IgG IP sample lanes.

Article Snippet: Reciprocal co-IPs were performed on whole rat retina lysate to confirm identified Tulp1-binding partners using the following target antibodies: MAP1B (NB100-68256, Novus Biologicals), Kif3a (EPR5087, Abcam, Cambridge, MA, USA), and Ribeye (612044, BD Transduction, San Francisco, CA, USA).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Photoreceptor Compartment-Specific TULP1 Interactomes

doi: 10.3390/ijms22158066

Figure Lengend Snippet: Immunolocalization of MAP1B and Tulp1 in P17 mouse retinas. ( A ) Wt retinal sections stained with MAP1B (red) and Tulp1 (green). ( B ) Tulp1−/− retinal sections stained with MAP1B (red) and Tulp1 (green). Sections were counterstained with DAPI (blue). Scale bar: 50 µm. INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS, inner segment layer; OS, outer segment layer.

Article Snippet: Reciprocal co-IPs were performed on whole rat retina lysate to confirm identified Tulp1-binding partners using the following target antibodies: MAP1B (NB100-68256, Novus Biologicals), Kif3a (EPR5087, Abcam, Cambridge, MA, USA), and Ribeye (612044, BD Transduction, San Francisco, CA, USA).

Techniques: Staining