Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mechanical Stress Induced NOX2 Promotes Endothelial Dysfunction in Ventilator-Induced Lung Injury: Potential Treatment with Quercetin.

doi: 10.1002/advs.202502639

Figure Lengend Snippet: Figure 3. Elevated ROS induce CaMKII/ERK1/2 signaling pathway activation in mechanical ventilation-induced lung injury (VILI). A) Venn diagram of shared genes in two sets (GSE226807 and GSE114132) of differentially expressed genes in lungs from sham control and MV. B,C) Bubble plot of Gene Ontology (GO) enrichment analysis for 884 genes differentially expressed in both sets. The x-axis represents the Rich Factor, indicating the degree of enrichment. The color scale indicates the -log2 (FDR), reflecting the significance of enrichment. Biological process (B) and Cellular component (C). D) Bubble plot of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis for 884 genes differentially expressed in both sets. The x-axis represents the Rich Factor, indicating the degree of enrichment. The color scale indicates the -log2 (Q-value), reflecting the significance of enrichment. The differentially expressed genes between H-MV and control groups E). PCA showed the distribution of the samples in the control group and the H-MV group F). The downregulated and unregulated genes in H-MV group compared to control group G). The volcano plot visually displays the distribution of differential genes H). Gene Ontology (GO) analysis between H-MV group and control group I). KEGG enrichment analysis between H-MV group and control group J). K) IPA predicts the signaling regulatory network associated with endothelial function, in which the NOX2/ROS/CaMKII/ERK1/2 signaling axis is highlighted in red. L,M) Representative Western blot (L) and the quantitative analysis (M) of phosphorylations and total expressions of CaMKII and ERK1/2 in lung tissue homogenates from the Control, L-MV, and H-MV groups respectively. Data are presented as the fold changes relative to the Control group. n = 6. N) Correlation analysis of p-CaMKII expression and DHE mean intensity defined ROS levels indicating that ROS level is positive correlated with p-CaMKII expression. R = 0.5973, p = 0.0187. n = 15. O) Correlation analysis of p- ERK1/2 expression and DHE mean intensity defined ROS levels indicating that ROS level is positive correlated with ERK1/2 expression. R = 0.7000, p = 0.0048. n = 15. P,Q) Representative Western blot (P) and the quantitative analysis (Q) of phosphorylations of CaMKII and ERK1/2 in HAECs stimulated with H2O2 at concentrations of 0 μM, 100 μM, 200 μM, and 500 μM respectively. Data are presented as the fold changes relative to the 0 nM group. n = 3. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. L-MV: low mechanical ventilation; H-MV: high mechanical ventilation; P-CaMKII: phosphorylated CaMKII; P-ERK1/2: phosphorylated ERK1/2; H2O2: hydrogen peroxide.

Article Snippet: The following primary antibodies were used: NOX1 (1:1000, Cat#: NBP1-31546, Novus), NOX2 (1:1000, Cat#: NBP2-41291, Novus), NOX4 (1:1000, Cat#: NB110-58849, Novus), Zonula occludin 1 (ZO-1) (1:1000, Cat#: 61–7300, Invitrogen), Occludin (1:1000, Cat#: 33–1500, Invitrogen), VE-cadherin (1:1000, Cat#: V1514-200UL, Sigma-Aldrich); P-CaMKII (1:1000, Cat#: SC-32289, Santa Cruz Biotechnology Inc), CaMKII (1:1000, Cat#: SC-5306, Santa Cruz Biotechnology Inc), P-ERK1/2 (1:1000, Cat#: 4370, Cell Signaling Technology, Danvers, MA, USA), ERK1/2 (1:1000, Cat#: 4695, Cell Signaling Technology), GAPDH(1:2000, Cat#: 60004-1-Ig, Proteintech), β-actin(1;2000, Cat#: 2876, Proteintech).

Techniques: Activation Assay, Control, Western Blot, Expressing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mechanical Stress Induced NOX2 Promotes Endothelial Dysfunction in Ventilator-Induced Lung Injury: Potential Treatment with Quercetin.

doi: 10.1002/advs.202502639

Figure Lengend Snippet: Figure 4. High cyclic stretch disrupts endothelial junctions via NOX2/ROS/CaMKII/ERK1/2 Axis. A,B) Representative DHE fluorescent images (A) and quantitative analysis (B) in control or H2O2 treated HAECs co-incubated with or without KN93 (a selective inhibitor for CaMKII). Scale bars: 30 μm. n = 6. C,D) Representative Western blot (C) and the quantitative analysis (D) of phosphorylations of CaMKII and ERK1/2 in control or H2O2 treated HAECs co-incubated with or without KN93. Data are presented as the fold changes relative to the Control group. n = 4. E–G) Representative immunofluorescent images of VE-cadherin (E) and ZO-1 (F) and the quantitative analysis (G) of ZO-1 and VE-cadherin in control or H2O2 treated HAECs co-incubated with or without KN93. Nuclei are counterstained with DAPI. Scale bars: 20 μm. The fluorescence intensity of ZO-1 and VE-cadherin is quantified using Image J software. n = 5–6. H,I) Representative Western blot (H) and the quantitative analysis (I) of phosphorylations of CaMKII and ERK1/2 in HAECs pre- treated with or without KN93 under static or CS conditions. Data are presented as the fold changes relative to the static group. n = 4. J,K) Representative Western blot (J) and the quantitative analysis (K) of phosphorylations of CaMKII and ERK1/2 in HAECs transfected with siNC or siNOX2 under static and CS conditions. Data are presented as the fold changes relative to the ST + siNC group. n = 4. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. H2O2: hydrogen peroxide; P-CaMKII: phosphorylated CaMKII; P-ERK1/2: phosphorylated ERK1/2; VE-cad: VE-cadherin; DAPI: 4′,6-diamidino-2-phenylindole; ST: static; CS: cyclic stretch; NC: negative control.

Article Snippet: The following primary antibodies were used: NOX1 (1:1000, Cat#: NBP1-31546, Novus), NOX2 (1:1000, Cat#: NBP2-41291, Novus), NOX4 (1:1000, Cat#: NB110-58849, Novus), Zonula occludin 1 (ZO-1) (1:1000, Cat#: 61–7300, Invitrogen), Occludin (1:1000, Cat#: 33–1500, Invitrogen), VE-cadherin (1:1000, Cat#: V1514-200UL, Sigma-Aldrich); P-CaMKII (1:1000, Cat#: SC-32289, Santa Cruz Biotechnology Inc), CaMKII (1:1000, Cat#: SC-5306, Santa Cruz Biotechnology Inc), P-ERK1/2 (1:1000, Cat#: 4370, Cell Signaling Technology, Danvers, MA, USA), ERK1/2 (1:1000, Cat#: 4695, Cell Signaling Technology), GAPDH(1:2000, Cat#: 60004-1-Ig, Proteintech), β-actin(1;2000, Cat#: 2876, Proteintech).

Techniques: Control, Incubation, Western Blot, Software, Transfection, Negative Control

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mechanical Stress Induced NOX2 Promotes Endothelial Dysfunction in Ventilator-Induced Lung Injury: Potential Treatment with Quercetin.

doi: 10.1002/advs.202502639

Figure Lengend Snippet: Figure 5. Quercetin effectively prevents mechanical stretch-induced endothelial dysfunction by scavenging ROS in vitro. A–C) Representative immunoflu- orescent images of ZO-1 (A) and VE-cadherin (B) and the quantitative analysis (C) in control or H2O2 treated HAECs co-incubated with or without quercetin. Nuclei are counterstained with DAPI. Scale bars: 20 μm. The fluorescence intensity of ZO-1 and VE-cadherin is quantified using Image J software. n = 6. Representative Western blot and the quantitative analysis of NOX2 (D). n = 4. E,F) Representative Western blot (E) and the quantitative analysis (F) of phosphorylations of CaMKII and ERK1/2 in HAECs pre-treated with or without quercetin under static or CS. Data are presented as the fold changes relative to the static group. n = 4. G,H) Representative Western blot (G) and the quantitative analysis (H) of protein expressions of ZO-1 and VE-cadherin in HAECs pre-treated with or without quercetin under static or CS. Data are presented as the fold changes relative to the static group. n = 4. I–K) Representative immunofluorescent images of ZO-1 (I) and VE-cadherin (J) and the quantitative analysis (K) in HAECs pre-incubated with or without quercetin under static or CS. Nuclei are counterstained with DAPI. Scale bars: 20 μm. The fluorescence intensity of ZO-1 and VE-cadherin is quantified using Image J software. n = 5-6. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. L-MV: low mechanical ventilation; H-MV: high mechanical ventilation; P-CaMKII: phosphorylated CaMKII; P-ERK1/2: phosphorylated ERK1/2; VE-cad: VE-cadherin; H2O2: hydrogen peroxide; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: The following primary antibodies were used: NOX1 (1:1000, Cat#: NBP1-31546, Novus), NOX2 (1:1000, Cat#: NBP2-41291, Novus), NOX4 (1:1000, Cat#: NB110-58849, Novus), Zonula occludin 1 (ZO-1) (1:1000, Cat#: 61–7300, Invitrogen), Occludin (1:1000, Cat#: 33–1500, Invitrogen), VE-cadherin (1:1000, Cat#: V1514-200UL, Sigma-Aldrich); P-CaMKII (1:1000, Cat#: SC-32289, Santa Cruz Biotechnology Inc), CaMKII (1:1000, Cat#: SC-5306, Santa Cruz Biotechnology Inc), P-ERK1/2 (1:1000, Cat#: 4370, Cell Signaling Technology, Danvers, MA, USA), ERK1/2 (1:1000, Cat#: 4695, Cell Signaling Technology), GAPDH(1:2000, Cat#: 60004-1-Ig, Proteintech), β-actin(1;2000, Cat#: 2876, Proteintech).

Techniques: In Vitro, Control, Incubation, Software, Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mechanical Stress Induced NOX2 Promotes Endothelial Dysfunction in Ventilator-Induced Lung Injury: Potential Treatment with Quercetin.

doi: 10.1002/advs.202502639

Figure Lengend Snippet: Figure 6. Antioxidant drug quercetin effectively attenuates ventilation-induced lung injury (VILI). A. Representative Western blot, and the quantitative analysis of NOX2 in lung tissue homogenates from the H-MV groups pre-treated with or without quercetin respectively. n = 5. B,C) Representative Western blot (B), and the quantitative analysis (C) of phosphorylations and total expressions of CaMKII and ERK1/2 in lung tissue homogenates from the Control, L-MV, and H-MV groups pre-treated with or without quercetin respectively. Data are presented as the fold changes relative to the Control group. n = 6. D,E) Representative H&E images of mice lung tissue sections from the Control, L-MV, and H-MV groups pre-treated with or without quercetin (D), and the quantitative analysis of lung injury histological scores (E). Scale bars: 50 μm, n = 4–5. F,G) Representative Western blot (F), and the quantitative analysis (G) of protein expressions of ZO-1, VE-cadherin and Occludin in lung tissue homogenates from the Control, L-MV, and H-MV groups pre-treated with or without quercetin. Data are presented as the fold changes relative to the Control group. n = 6. H–K) Representative immunofluorescent images of ZO-1 (H), VE-cadherin (I), and Occludin (J) and the quantitative analysis (K) of ZO-1, VE-cadherin, and Occludin in mice lung tissue sections from the Control, L-MV and H-MV groups pretreated with or without quercetin. Nuclei are counterstained with DAPI. The average fluorescent intensity of ZO-1, Occludin and VE-adherin is quantified using Image J software. Scale bars: 20 μm, n = 6. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. L-MV: low mechanical ventilation; H-MV: high mechanical ventilation; P-CaMKII: phosphorylated CaMKII; P-ERK1/2: phosphorylated ERK1/2; VE-cad: VE-cadherin; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: The following primary antibodies were used: NOX1 (1:1000, Cat#: NBP1-31546, Novus), NOX2 (1:1000, Cat#: NBP2-41291, Novus), NOX4 (1:1000, Cat#: NB110-58849, Novus), Zonula occludin 1 (ZO-1) (1:1000, Cat#: 61–7300, Invitrogen), Occludin (1:1000, Cat#: 33–1500, Invitrogen), VE-cadherin (1:1000, Cat#: V1514-200UL, Sigma-Aldrich); P-CaMKII (1:1000, Cat#: SC-32289, Santa Cruz Biotechnology Inc), CaMKII (1:1000, Cat#: SC-5306, Santa Cruz Biotechnology Inc), P-ERK1/2 (1:1000, Cat#: 4370, Cell Signaling Technology, Danvers, MA, USA), ERK1/2 (1:1000, Cat#: 4695, Cell Signaling Technology), GAPDH(1:2000, Cat#: 60004-1-Ig, Proteintech), β-actin(1;2000, Cat#: 2876, Proteintech).

Techniques: Western Blot, Control, Software

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Mechanical Stress Induced NOX2 Promotes Endothelial Dysfunction in Ventilator-Induced Lung Injury: Potential Treatment with Quercetin.

doi: 10.1002/advs.202502639

Figure Lengend Snippet: Figure 7. Quercetin markedly improves survival and alleviates acute lung injury in cecal ligation puncture (CLP)-induced septic mice. A) Venn diagram of shared genes between 884 genes in Figure 3. A and the differentially expressed genes in lungs between normal control and CLP (GSE226807). B,C) Bubble plot of Gene Ontology (GO) enrichment analysis for 796 genes differentially expressed in both sets. The x-axis represents the Rich Factor, indicating the degree of enrichment. The color scale indicates the -log2 (FDR), reflecting the significance of enrichment. D) Bubble plot of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis for 796 genes differentially expressed in both sets. The x-axis represents the Rich Factor, indicating the degree of enrichment. The color scale indicates the -log2 (Q-value), reflecting the significance of enrichment. E,F) Survival curves of male (E) and female (F) mice pre-treated with or without quercetin in the sham and CLP groups. G) Quantitative analysis of lung wet-to-dry weight ratio in the Control and CLP groups. n = 6. Representative Western blot (H) and quantitative analysis of NOX2 (I) from CLP model pretreated with or without quercetin. n = 5. J,K) Representative Western blot images (J) and quantitative analysis (K) of phosphorylations of CaMKII and ERK1/2 in lung tissue homogenates from the Control and CLP groups pretreated with or without quercetin. Data are presented as the fold changes relative to the shame group. n = 5. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. P-CaMKII: phosphorylated CaMKII; P-ERK1/2: phosphorylated ERK1/2; CLP: cecal ligation puncture.

Article Snippet: The following primary antibodies were used: NOX1 (1:1000, Cat#: NBP1-31546, Novus), NOX2 (1:1000, Cat#: NBP2-41291, Novus), NOX4 (1:1000, Cat#: NB110-58849, Novus), Zonula occludin 1 (ZO-1) (1:1000, Cat#: 61–7300, Invitrogen), Occludin (1:1000, Cat#: 33–1500, Invitrogen), VE-cadherin (1:1000, Cat#: V1514-200UL, Sigma-Aldrich); P-CaMKII (1:1000, Cat#: SC-32289, Santa Cruz Biotechnology Inc), CaMKII (1:1000, Cat#: SC-5306, Santa Cruz Biotechnology Inc), P-ERK1/2 (1:1000, Cat#: 4370, Cell Signaling Technology, Danvers, MA, USA), ERK1/2 (1:1000, Cat#: 4695, Cell Signaling Technology), GAPDH(1:2000, Cat#: 60004-1-Ig, Proteintech), β-actin(1;2000, Cat#: 2876, Proteintech).

Techniques: Ligation, Control, Western Blot

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: CAMK2B expression in pan carcinoma and survival analysis of CAMK2B in patients with glioma. ( A ) CAMK2B expression in pan carcinoma. ( B ) Survival analysis of patients from the Second Hospital of Hebei Medical University. ( C ) Survival analysis of patients from TCGA, CCGA, and Rambrandt databases

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Expressing

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: CAMK2B expression in tissues and cells. ( A ) Immunohistochemical results showed that compared with normal brain tissue, CAMK2B expression level in glioma decreased with the increase of glioma grade. ( B , C ) qRT-PCR and Western Blot results show that CAMK2B is significantly low expressed in three glioma cells (U251, U87, A172) compared with human astrocytes (HA). * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Expressing, Immunohistochemical staining, Quantitative RT-PCR, Western Blot

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: Stimulation of CAMK2B inhibits the proliferation of glioma cells. ( A , B ) Detection of CAMK2B expression in U251 cells after plasmid transfection by qRT-PCR and Western Blot. ( C ) CCK-8 assay reveals the effect of CAMK2B on U251 and U87 glioma cell viability. ( D ) EdU assay reveals the effect of CAMK2B on U251 and U87 glioma cell proliferation. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Expressing, Plasmid Preparation, Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, EdU Assay

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: Activating CAMK2B inhibits the invasion and migration of glioma cells. ( A ) The Transwell assay displayed that activated CAMK2B reduced the invasion of U251 and U87 glioma cells. ( B ) The wound healing assay displayed that activated CAMK2B reduced the healing area of U251 and U87 glioma cells. * p < 0.05; *** p < 0.001; **** p < 0.0001

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Migration, Transwell Assay, Wound Healing Assay

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: Silencing CAMK2B can promote the proliferation of glioma cells in vivo . ( A , B ) Detection of CAMK2B expression after siCAMK2B transfection by qRT-PCR and Western Blot. ( C , D ) Detection of CAMK1D and CAMK2A expression after siCAMK2B transfection by qRT-PCR. ( E , F ) Tumor volume and weight in vivo were significantly increased after subcutaneous injection of U251 cells of siCAMK2B into nude mice. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; “ns” stands for no statistical significance

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: In Vivo, Expressing, Transfection, Quantitative RT-PCR, Western Blot, Injection

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: CAMK2B expression affects Ras/Raf/MEK/ERK signal pathway. ( A ) The Ras, p-Raf, p-MEK, and p-ERK were increased after the knock-down of CAMK2B; nevertheless, the Ras inhibitor Salirasib can reverse this change. ( B ) The Ras, p-Raf, p-MEK, and p-ERK were decreased after activating CAMK2B

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Expressing, Knockdown

Journal: Oncology Research

Article Title: CAMK2B Impacts the Proliferation, Invasion, and Migration of Glioma Cells via the Ras/Raf/MEK/ERK Signaling Pathway

doi: 10.32604/or.2025.064300

Figure Lengend Snippet: Silencing CAMK2B can promote the proliferation, invasion, and migration of glioma cells, while the Ras inhibitor Salirasib can reverse this effect. ( A ) CCK8 assay points out that silencing CAMK2B can enhance the viability of U251 and U87 glioma cells, and the Ras inhibitor Salirasib can reverse this result. ( B ) EdU assay displayed that silencing CAMK2B could enhance the proliferation of U251 and U87 glioma cells; simultaneously, the Ras inhibitor Salirasib could reverse this result. ( C ) Transwell assay shows that knockdown of CAMK2B can improve the invasive ability of U251 and U87 glioma cells; concurrently, the Ras inhibitor Salirasib can reverse this trend. ( D ) Wound healing assay indicated that the knockdown of CAMK2B can raise the migration ability of U251 and U87 glioma cells, besides the Ras inhibitor Salirasib can reverse this phenomenon. * p < 0.05; ** p < 0.01; **** p < 0.0001; “ns” stands for no statistical significance

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) (SB-PR079, ShareBio, Shanghai, China) at room temperature for 40 min, and incubated overnight at 4°C with primary antibodies against CAMK2B (1:1000) (11533-1-AP, Proteintech, Rosemont, IL, USA), GAPDH (1:7500) (10494-1-AP, Proteintec), Ras (1:1000) (12063-1-AP, Proteintech), Raf1 (1:1000) (26863-1-AP, Proteintech).

Techniques: Migration, CCK-8 Assay, EdU Assay, Transwell Assay, Knockdown, Wound Healing Assay