Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 1 ITGBL1 is upregulated in PCa tissues. (A) In the mRNA sequencing of PCa dataset from TCGA, ITGBL1 expression levels were upregulated in PCa tissues compared with the normal adjacent tissues (ANT). PCa: n=497; ANT=52. (B) ITGBL1 expression levels were upregulated in 52 paired PCa tissues compared with the matched normal adjacent tissues as assessed by the mRNA sequencing ofPCa dataset from TCGA. (C) The real-time PCR analysis of ITGBL1 expression in 8 paired PCa samples. Transcript levels were normalized to Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. Error bars represent the mean ± S.D. of three independent experiments. *P<0.05. (D) Western blot analysis of ITGBL1 expression in 8 paired PCa samples. α-Tubulin served as the loading control. (E) The real- time PCR analysis of ITGBL1 expression levels in the normal prostate epithelial cell (RWPE-1), primary PCa cell 22RV1, bone metastatic PCa cell lines (PC-3 and VCaP), brain metastatic cell line DU145 and lymph node metastatic cell line LNCaP. Transcript levels were normalized to GAPDH expression. Error bars represent the mean ± S.D. of three independent experiments. *P<0.05. (F) Western blot analysis of ITGBL1 expression in a normal prostate epithelial cell (RWPE-1), primary PCa cell 22RV1, bone metastatic PCa cell lines (PC-3 and VCaP), brain metastatic cell line DU145 and lymph node metastatic cell line LNCaP. α-Tubulin served as the loading control. Abbreviations: TCGA, the cancer genome atlas; ANT ,the normal adjacent tissues;GAPDH,glyceradehyde-3-phosphate dehydrogenase;ITGBL1, integrin beta-like1.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Sequencing, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 2 Overexpression of ITGBL1 is associated with lymph node metastasis of PCa. (A) Representative images of ITGBL1 expression in PCa tissues with lymph node metastasis and PCa tissues without lymph node metastasis via immunohistochemistry (IHC). (B) The IHC staining index of ITGBL1 expression in PCa tissues. (C) Percentages and number of samples showed high or low ITGBL1 expression in our PCa patients with different Gleason scores. (D) Percentages and number of samples showed high or low ITGBL1 expression in our PCa patients with different tumor size. (E) Percentages and number of samples showed high or low ITGBL1 expression in our PCa patients with different lymph node metastasis. (F) Percentages and number of samples showed high or low ITGBL1 expression in our PCa patients with different distant metastasis. (G) Percentages and number of samples showed high or low ITGBL1 expression in PCa tissues with different Gleason scores from TCGA. *P<0.05. (H) Kaplan- Meier progression-free survival curves for PCa patients stratified by high and low expression of ITGBL1 as assessed by analyzing the RNA sequencing dataset of PCa from TCGA. (I) Kaplan-Meier overall survival curves for PCa patients stratified by high and low expression of ITGBL1 as assessed by analyzing the RNA sequencing dataset of PCa from TCGA. Abbreviations: IHC, immunohistochemistry; ITGBL1, integrin beta-like1; TCGA, the cancer genome atlas.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Over Expression, Expressing, Immunohistochemistry, RNA Sequencing

Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 3 ITGBL1 promotes EMT in PCa cells. (A–D) Gene set enrichment analysis (GSEA) revealed that ITGBL1 expression significantly and positively correlated with the EMT signatures and metastatic phenotypes. (E) The real-time PCR analysis of ITGBL1 expression in the indicated PCa cells compared to the corresponding controls. Transcript levels were normalized by GAPDH expression. *P<0.05. (F) Western blot analysis of ITGBL1 expression in the indicated PCa cells compared to the corresponding controls. α-Tubulin served as the loading control. (G) Upregulation of ITGBL1 converted a cobblestone-like or a short spindle-shaped epithelial morphology to a stick-like or long spindle-shaped mesenchymal profile in 22RV1 cells. Conversely, silencing ITGBL1 converted a stick-like or long spindle-shaped mesenchymal profile to a cobblestone-like or a short spindle-shaped epithelial morphology in LNCaP cells. (H) Upregulation of ITGBL1 decreased E-cadherin expression and increased Vimentin and Fibronectin expression in 22RV1 cells while silencing ITGBL1 increased E-cadherin expression and decreased Vimentin and Fibronectin expression in LNCaP cells. α- Tubulin served as the loading control.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 4 ITGBL1 promotes invasion and migration abilities in PCa cells. (A and B) Invasion assay indicated upregulating ITGBL1 enhanced, while silencing ITGBL1 reduced the invasion and migration ability of PCa cells. Histograms show the relative invasion ability in the indicated cells.*P<0.05.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Migration, Invasion Assay

Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 5 ITGBL1 promotes invasion and migration ability via activating NF-κB signaling pathway in PCa cells. (A and B) GSEA revealed that ITGBL1 expression was positively associated with NF-κB signaling. (C and D) NF-κB transcriptional activity was assessed by luciferase reporter constructs in the indicated PCa cells. Error bars represent the mean ± S.D. of three independent experiments. *P<0.05. (E) Western blotting of nuclear NF-κB/p65 expression in the indicated PCa cells. The nuclear protein p84 was used as the nuclear protein marker, respectively.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Migration, Expressing, Activity Assay, Luciferase, Construct, Western Blot, Marker

Journal: OncoTargets and Therapy

Article Title:

ITGBL1 promotes EMT, invasion and migration by activating NF-κB signaling pathway in prostate cancer

doi: 10.2147/ott.s200082

Figure Lengend Snippet: Figure 6 The activity of NF-kB signaling is essential for ITGBL1-induced invasion and migration, and tumorigenesis of PCa cells. (A) NF-κB signaling inhibitors LY2409881 and JSH-23 decreased the stimulatory effect of ITGBL1 on NF-κB transcriptional activity in PCa cells respectively. Error bars represent the mean ± s.d. of three independent experiments. *P<0.05. The NFκB luciferase activity results presented in (A) were independent experiments. (B) NF-κB signaling inhibitors LY2409881 and JSH-23 attenuated the stimulatory effects of ITGBL1 on invasion and migration abilities of PCa cells. Error bars represent the mean ± s.d. of three independent experiments. *P<0.05. (C) Representative images of the tumors are shown in the xenograft model of nude mice. (D) Tumor volumes in the indicated groups were measured every 4 days. Data presented are the mean ± s.d. *P<0.05. (E) Tumor weights in the tumor tissue from the indicated mice groups. *P<0.05.

Article Snippet: The control and pNFκB-luc plasmids were purchased from Promega Corporation. shRNAs or plasmids were transfected using Lipofectamine 3000 (Life Technologies) as previously described.13 Western blotting Western blotting was performed as described previously.14 The E-cadherin (Cat#. 3195), Vimentin (Cat#. 5741) and Fibronectin (Cat#. 4706) antibodies were purchased from Cell Signaling Technology, ITGBL1 (Cat#: Ag11521) and p65 (Cat#. 10745–1-AP) from Proteintech, p84 (Cat#. PA5–27816) from Invitrogen.

Techniques: Activity Assay, Migration, Luciferase

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Exercise preconditioning prevented muscle atrophy through protein arginine methyltransferase 1 (Prmt1)-Sestrin1 (Sesn1)-transcriptional co-activator PPAR-γ co-activator-1 α (PGC-1α)-mediated skeletal muscle regeneration. (A–B) Western blot results of MyoD, Myf5, MEF2 and MyoG in GAS muscle. (C–D) Co-IP results, IP: MyoD, GAS muscle was used. A-B, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used, and data are shown as means ​± ​standard error of the mean ( SEM ) (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8). (E–F) H&E staining of myotubes at each differentiation time points of C2C12 myoblasts overexpressing Sesn1. Scale bar ​= ​100 ​μm ∗ p ​< ​0.05 vs. Ad-Sesn1-, unpaired Student's t-test was used and data are shown as means ​± ​ SEM ( n ​= ​3 in each group). (G–H) Western blot results of Sesn1, Prmt1, PGC-1α, MyoD, MEF2, Myf5, and MyoG in C2C12 myoblasts at each time points of differentiation. ∗ p ​< ​0.05 vs. D0, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (I–J) Western blot results of PGC-1α in nucleus of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (K–L) Western blot results of PGC-1α in cytoplasm of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1-, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group).

Article Snippet: The antibodies are listed below: aDMA (Anti-Asymmetric Di-Methyl Arginine Motif) (1:1 500, Rabbit, Cell Signal Tech, USA), Akt (protein kinase B) (1:2 000, Mouse, Proteintech, USA), pAkt-Ser473 (1:2 000, Rabbit, Cell Signal Tech, USA), AMPKα2 (1:2 000, Rabbit, Cell Signal Tech, USA), pAMPK-Thr172 (1:2 000, Rabbit, Cell Signal Tech, USA), ATF2 (1:2 000, Rabbit, Proteintech, USA), Atrogin-1 (FBXO32) (1:15 000, Mouse, Proteintech, USA), FoxO3a (Forkhead box O3) (1:1 000, Mouse, Proteintech, USA), pFoxO3a-Ser315 (1:2 000, Rabbit, Proteintech, USA), GAPDH (1:5 000, Rabbit, Utibody, CN), IGF-1 (insulin-like growth factor 1) (1:2 000, Mouse, Proteintech, USA), LaminB (1:2 000, Rabbit, Abcam, USA), MEF2 (myocyte enhancer factor 2) (1:2 000, Rabbit, Proteintech, USA), Myf5 (myogenic factor 5) (1:2 000, Rabbit, Abclonal, CN), MyoD (myogenic differentiation antigen) (1:2 000, Rabbit, Proteintech, USA), MyoG (myogenin) (1:2 000, Rabbit, Proteintech, USA). mTOR (mammalian target of rapamycin) (1:2 000, Rabbit, Cell Signal Tech, USA), p38 (1:2 000, Rabbit, WANLEIBIO, CN), p-p38-Thr180/Tyr182 (1:2 000, Rabbit, WANLEIBIO, CN), PGC-1α (1:1 000, Rabbit, Abcam, USA), Prmt1 (1:2 000, Rabbit, Cell Signal Tech, USA), Raptor (1:1 000, Rabbit, Cell Signal Tech, USA), Sesn1 (1:1 000, Rabbit, HUABIO, CN), Sesn1 (1:1 000, Rabbit, Abcam, USA), TRIM63 (MuRF1) (1:2 000, Rabbit, Proteintech, USA).

Techniques: Western Blot, Co-Immunoprecipitation Assay, Staining

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B increases HO-1 and TREM2 expression in perihematomal brain tissue and microglia after ICH. ( A – C ) Immunoblotting showing relative HO-1 and TREM2 protein expression in microglia under the different treatments ( n = 4). ( D , E ) Quantitative PCR (qRT-PCR) assessment of HO-1 and TREM2 transcript levels in microglia under the different treatments ( n = 3). ( F – H ) Protein analysis by Western blot for HO-1 and TREM2 in perihematomal tissue from ICH mice on post-ICH day 3 under the different treatments ( n = 4) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B upregulates HO-1 and TREM2 expression via Nrf2. ( A , B ) Immunoblotting for relative Nrf2 protein expression in microglia under the different treatments ( n = 3). ( C ) Quantitative PCR (qRT-PCR) assessment of relative Nrf2 transcript levels in microglia under the different treatments ( n = 3). ( D ) Confocal micrographs depicting Nrf2 expression in microglia under the different treatments ( n = 3; scale bar = 10 μm). ( E – G ) Protein analysis by Western blot for HO-1 and TREM2 in microglia under the different treatments ( n = 3) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B promotes hematoma absorption after ICH by enhancing PlexinA1-mediated microglial phagocytic function. Exogenous supplementation of Sema3B binds to its receptor PlexinA1, activating the DAP12-dependent signaling pathway (Syk-PI3K-AKT-mTOR) and NRF2 in microglia, thereby increasing the expression of TREM2 and HO-1 to facilitate microglia-mediated hematoma clearance, while suppressing neuroinflammation by inhibiting the NF-κB pathway and reducing pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). This figure was drawn by Figdraw.

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing