Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: Immunochemistry data confirmed the presence of BEX1 and MAP1b in various cell types. (A) Violin plot showing the expression of BEX1 and MAP1b in each cell type. (B) Violin plot showing the expression of BEX1 and MAP1b in tumor tissue compared with adjacent noncancerous tissue. (C,D) Representative immunohistochemical staining of BEX1 (C) and MAP1b (D) in tumor tissue and adjacent noncancerous tissue. *, P<0.05; ***, P<0.001. N, noncancerous tissue; NK, natural killer; T, tumor tissue.

Article Snippet: Slides were subsequently treated at 4 °C overnight with an antibody against BEX1 (Proteintech, Rosemont, IL, USA; Cat#12330-1-AP, 1:100) and an antibody against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666, 1:400).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: MAP1b and BEX1 modulated proliferation, migration and apoptosis in vitro. (A) Levels of MAP1b and BEX1 mRNA in different SCLC cell lines. (B,C) RT-qPCR and Western blotting were applied to examine the knockdown efficiency of MAP1b in NCI-H82 and BEX1 in NCI-H209. (D) A CCK-8 assay was conducted to detect cell proliferation following MAP1b knockdown (left) and BEX1 knockdown (right) in NCI-H82 and NCI-H209. (E) The effect of MAP1b (left) and BEX1 (right) knockdown on the migration capacity of NCI-H82 and NCI-H209. (F-I) The effect of MAP1b (F,G) and BEX1 (H,I) knockdown on cell apoptosis in NCI-H82 and NCI-H209, respectively. Unpaired two-tailed Student’s t -tests and one-way or two-way analysis of variance were used to determine significance. Data are presented as mean ± SD (n=3). **, P<0.01; ****, P<0.0001. CCK-8, Cell Counting Kit-8; OD, optical density; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SCLC, small cell lung cancer; SD, standard deviation.

Article Snippet: Slides were subsequently treated at 4 °C overnight with an antibody against BEX1 (Proteintech, Rosemont, IL, USA; Cat#12330-1-AP, 1:100) and an antibody against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666, 1:400).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Western Blot, Knockdown, CCK-8 Assay, Two Tailed Test, Cell Counting, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: Immunochemistry data confirmed the presence of BEX1 and MAP1b in various cell types. (A) Violin plot showing the expression of BEX1 and MAP1b in each cell type. (B) Violin plot showing the expression of BEX1 and MAP1b in tumor tissue compared with adjacent noncancerous tissue. (C,D) Representative immunohistochemical staining of BEX1 (C) and MAP1b (D) in tumor tissue and adjacent noncancerous tissue. *, P<0.05; ***, P<0.001. N, noncancerous tissue; NK, natural killer; T, tumor tissue.

Article Snippet: Membranes were incubated on a plate shaker overnight at 4 °C with corresponding primary antibodies against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666), BEX1 (ThermoFisher Cat#PA5-100404, Thermo Fisher Scientific, Waltham, MA, USA), and Actin (Abcam Cat#ab8226, RRID: AB_306371, Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: MAP1b and BEX1 modulated proliferation, migration and apoptosis in vitro. (A) Levels of MAP1b and BEX1 mRNA in different SCLC cell lines. (B,C) RT-qPCR and Western blotting were applied to examine the knockdown efficiency of MAP1b in NCI-H82 and BEX1 in NCI-H209. (D) A CCK-8 assay was conducted to detect cell proliferation following MAP1b knockdown (left) and BEX1 knockdown (right) in NCI-H82 and NCI-H209. (E) The effect of MAP1b (left) and BEX1 (right) knockdown on the migration capacity of NCI-H82 and NCI-H209. (F-I) The effect of MAP1b (F,G) and BEX1 (H,I) knockdown on cell apoptosis in NCI-H82 and NCI-H209, respectively. Unpaired two-tailed Student’s t -tests and one-way or two-way analysis of variance were used to determine significance. Data are presented as mean ± SD (n=3). **, P<0.01; ****, P<0.0001. CCK-8, Cell Counting Kit-8; OD, optical density; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SCLC, small cell lung cancer; SD, standard deviation.

Article Snippet: Membranes were incubated on a plate shaker overnight at 4 °C with corresponding primary antibodies against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666), BEX1 (ThermoFisher Cat#PA5-100404, Thermo Fisher Scientific, Waltham, MA, USA), and Actin (Abcam Cat#ab8226, RRID: AB_306371, Abcam, Cambridge, UK).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Western Blot, Knockdown, CCK-8 Assay, Two Tailed Test, Cell Counting, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

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