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: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: FAM65A binds to Ras and activates the Ras/ERK signaling to mediate RSK activation (A) The volcano plot analysis results for the FAM65A high-expression and low-expression groups from the TCGA database were shown. (B) The KEGG and GO results were shown. (C) The GSEA results were shown. (D) GSEA on DEGs between the FAM65A high-expression group and low-expression group in the Reactome database were shown. (E) IP was performed to detect the binding of FAM65A and Ras/p-RSK. (F) IP was performed to detect the binding of Ras and FAM65A/p-RSK. (G) Immunofluorescence was performed to detect the co-localization of FAM65A and Ras. Scale bars, 20 μm. (H) Western blot analysis the Ras and p -ERK expression in FAM65A knockdown or overexpression cells. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Expressing, Binding Assay, Immunofluorescence, Western Blot, Knockdown, Over Expression

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Ras/ERK signaling activation was indispensable for FAM65A-mediated RSK activation and CRC progression (A) Western blot analysis of Ras and p -ERK expression in HCT116-FAM65A cells treated with 10 μM Abd-7, or without treatment. (B) Results from the CCK8 cell proliferation assay conducted on HCT116-FAM65A cells with and without the application of Abd-7, n = 3, ∗∗∗ p < 0.001. (C) Colony formation assay performed on HCT116-FAM65A cells treated with Abd-7 or not. (D) Quantitative analysis of the colony formation assay results, n = 3, ∗∗∗ p < 0.001. (E) Results from the EdU assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 100 μm. (F) Quantitative analysis of the EdU assay results, n = 3, ∗∗∗ p < 0.001. (G) Western blot analysis of Ki-67, cleaved Caspase 3, Bcl-2, and Bax expression in HCT116-FAM65A cells treated with Abd-7 or not. (H) Results from the apoptosis assay conducted on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (I) Quantitative analysis of the apoptosis experiments, n = 3, ∗∗∗ p < 0.001. (J) Results from the Transwell migration assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 50 μm. (K) Quantitative analysis of the Transwell migration assay results, n = 3, ∗∗∗ p < 0.001. (L) Results from the wound healing assay performed on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (M) Quantitative analysis of the wound healing assay results, n = 3, ∗∗∗ p < 0.001. (N) Western blot analysis the expression of EMT markers in HCT116-FAM65A cells treated with Abd-7 or not. (O) Proposed model of FAM65A in CRC progression. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Western Blot, Expressing, Proliferation Assay, Colony Assay, EdU Assay, Apoptosis Assay, Transwell Migration Assay, Wound Healing Assay

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Knockdown of FAM65A inhibits tumor progression in vivo (A) LOVO-shCtrl and LOVO-shFAM65A cells were administered into the fourth fat pad of nude mice, and the resulting tumor growth curves were subsequently generated, n = 5, ∗ p < 0.05. (B) The tumors excised from mice across various experimental groups are presented. (C) Hematoxylin and Eosin (HE) staining results of lung tissue from the different groups is displayed. (D) A quantitative analysis of metastatic lung nodules is provided, n = 5, ∗∗ p < 0.01. (E) IHC results for FAM65A, Ki-67, p -RSK, p -ERK, Ras, N-cadherin, vimentin, cleaved Caspase 3, ZO-1, and E-cadherin in tumor tissues are illustrated. (F) A quantitative analysis of the IHC results is included. Data are presented as mean ± SEM of biologically independent experiments, n = 5, ∗∗∗ p < 0.001.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Knockdown, In Vivo, Generated, 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: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Validation of the targeting relationship between let-7a-5p and MAPK8. (A) Schematic representation of the predicted complementary binding site between let-7a-5p and the 3′-UTR of MAPK8. (B) Relative mRNA expression level of MAPK8 in macrophages overexpressing let-7a-5p detected by RT-qPCR. (C) WB analysis of MAPK8 protein expression in macrophages overexpressing let-7a-5p. (D) Inhibitory effect of let-7a-5p on MAPK8 expression was assessed via a dual luciferase reporter assay. ( **P < 0.01, ***P < 0.001).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Biomarker Discovery, Binding Assay, Expressing, Quantitative RT-PCR, Luciferase, Reporter Assay

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Effect of PRP on expression of let-7a-5p and MAPK8. (A) Relative mRNA expression levels of let-7a-5p in knee joint sections of each group detected by RT-qPCR. (B) iNOS and CD206 co-immunolabeld with MAPK8 and counter-stained with DAPI in synovial tissues (Scale bar: 50 μm). (C, D) Quantification of iNOS and CD206 expression co-localized with MAPK8. ( **P < 0.01).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: The let-7a-5p/MAPK8 axis regulates macrophage polarization and inflammatory cytokine release in vitro . (A) IF staining showing expression of iNOS and CD206 in macrophages (Scale bar: 50 μm). (B) Quantification of iNOS-positive cell rate in transfected macrophages. (C) Quantification of CD206-positive cell rate in transfected macrophages. (D-G) Relative mRNA expression levels of pro-inflammatory cytokine (IL-1β and TNF-α) and anti-inflammatory cytokine (IL-4 and IL-10) in transfected macrophages detected by RT-qPCR. ( *P < 0.05, **P < 0.01, ns no significance).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: In Vitro, Staining, Expressing, Transfection, Quantitative RT-PCR