Journal: Cancers

Article Title: Loss of PGRMC1 Delays the Progression of Hepatocellular Carcinoma via Suppression of Pro-Inflammatory Immune Responses

doi: 10.3390/cancers13102438

Figure Lengend Snippet: Loss of Pgrmc1 aggravates liver injury while suppressing compensatory proliferation. ( A ) WT and Pgrmc1 -null mice were injected with high-dose DEN (200 mg/kg, i.p.), and their plasma and livers were collected after 48 h. ( B ) Plasma level of alanine aminotransferase (ALT). ( C ) Expression of high mobility group box 1 (HMGB1) in the plasma. The expression level was normalized to that of plasma albumin and expressed relative to the WT group. ( D ) TUNEL staining in the livers of WT and Pgrmc1 -null mice. TUNEL-positive cells were counted and expressed relative to the WT group. Scale bar, 100 μm. ( E ) Ki67 immunostaining in the livers of WT and Pgrmc1 -null mice. Ki67-positive cells were counted and expressed relative to the WT group. Scale bar, 100 μm. ( F ) mRNA expression levels of C-Myc , Cyclin D , and Hgf in the livers of WT and Pgrmc1 -null mice. The expression level was normalized to that of Rplp0 and expressed relative to the WT group. * p < 0.05 in Student’s t -test. All full blot images were provided in .

Article Snippet: The following primary antibodies were used: β-actin (sc-130656, Santa Cruz Biotechnology), phosphor-IκBα (pIκBα), NF-κB p65, phosphor-NF-κB p65 (pNF-κB p65) (9936, CST, MA, USA), EGFR (A2909, ABclonal, MA, USA), phosphor-EGFR (9789, CST, MA, USA), HMGB1 (CSB-PA01604A0Rb, Cusabio, TX, USA), PGRMC1 (rabbit monoclonal, 13856, CST, MA, USA), and IκBα (mouse monoclonal, 9936, CST, MA, USA).

Techniques: Injection, Clinical Proteomics, Expressing, TUNEL Assay, Staining, Immunostaining

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Contains a summary of the Primer Sequence List.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Design and characterization of HMGB1-interfering lentiviruses (A) Schematic design of shRNA targeting HMGB1 mRNA. (B) Real-time PCR analysis of HMGB1 RNA levels in rat astrocytes infected with lentivirus expressing HMGB1 shRNA. (C) qRT-PCR analysis of HMGB1 gene expression in the spinal cord of rats injected with different titers. **** p < 0.0001 *** p < 0.001 ** p < 0.01* p < 0.05.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: shRNA, Real-time Polymerase Chain Reaction, Infection, Expressing, Quantitative RT-PCR, Gene Expression, Injection

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Mechanical and thermal pain threshold responses in CCD rats within each group (A) Mechanical pain thresholds in CCD rats after HMGB1 knockdown. (B) Thermal pain thresholds in CCD rats after HMGB1 knockdown. (C) Mechanical pain thresholds in CCD rats after TLR4 inhibition. (D) Thermal pain thresholds in CCD rats after TLR4 inhibition. (E) Mechanical pain thresholds in CCD rats after RAGE inhibition. (F) Thermal pain thresholds in CCD rats after RAGE inhibition. N = 5 per group **** p < 0.0001 *** p < 0.001 ** p < 0.01* p < 0.05 vs. CCD.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: Knockdown, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Immunoprecipitation determinations of HMGB1 and AQP1 (A) Western blots of HMGB1 in plasmids. (B) Western blots of AQP1 in plasmids. (C) CO-IP assay results. 293T: 293T-null cells; 293T-E5061-E5077: 293T- E5061 HA empty control plasmid transfection- E5077 negative control CON238 plasmid; 293T-E5062-E5078: 293T- E5062 HA-Aqp1 overexpression plasmid transfection- E5078 Hmgb1-3flag overexpression plasmid. Flag: HMGB1; HA:AQP1.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: Immunoprecipitation, Western Blot, Co-Immunoprecipitation Assay, Control, Plasmid Preparation, Transfection, Negative Control, Over Expression

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Changes in AQP1 expression after HMGB1 knockdown (A) Protein expression levels of HMGB1 and AQP1 in the spinal cords of rats in each group. (B) Protein expression levels of HMGB1 and AQP1 in the LPS inflammatory cell model in each group. (C) mRNA levels of HMGB1 and AQP1 in the spinal cords of rats in each group. (D) mRNA levels of HMGB1 and AQP1 in the LPS inflammatory cell model in each group. N = 3 per group **** p < 0.0001 *** p < 0.001 ** p < 0.01* p < 0.05.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: Expressing, Knockdown

Journal: Frontiers in Pharmacology

Article Title: Lentivirus-mediated RNA interference targeting HMGB1 modulates AQP1 to reduce pain induced by chronic compression of the dorsal root ganglia

doi: 10.3389/fphar.2024.1469223

Figure Lengend Snippet: Changes in NF-κB expression following knockdown of HMGB1 or AQP1 (A) Protein expression levels of HMGB1 and NF-κB in the spinal cords of rats in each group. (B) mRNA levels of HMGB1 and NF-κB in the spinal cords of rats in each group. (C) Protein expression levels of AQP1 and NF-κB in the spinal cords of rats in each group. (D) mRNA levels of AQP1 and NF-κB in the spinal cords of rats in each group. (E) Protein expression levels of NF-κB and AQP1 in the LPS inflammatory cell model in each group. (F) mRNA levels of NF-κB and AQP1 in the LPS inflammatory cell model in each group. N = 3 per group **** p < 0.0001 *** p < 0.001 ** p < 0.01* p < 0.05.

Article Snippet: The following primary antibodies were used: rabbit anti-HMGB1 antibody (20 μg, CUSABIO), rabbit anti-AQP1 antibody (20 μL, BOSTER BIOLOGICAL TECHNOLOGY), rabbit anti-RAGE antibody (10 μL, MedChemExpress), rabbit anti-GAPDH antibody (100 μL, BOSTER) and mouse anti-TLR4 antibody (20 μL, proteintech).

Techniques: Expressing, Knockdown