Journal: Cancers

Article Title: Short-Term TERT Inhibition Impairs Cellular Proliferation via a Telomere Length-Independent Mechanism and Can Be Exploited as a Potential Anticancer Approach

doi: 10.3390/cancers15102673

Figure Lengend Snippet: Telomerase reverse transcriptase (TERT) inhibition downregulated MYC proto-oncogene bHLH transcription factor (MYC) levels. ( A , B ) Cells were treated with BIBR at indicated concentrations or with DMSO as a control for 24 h. Levels of relative mRNA expression for MYC and TERT in 4134/Late ( A ) and BL41 ( B ) cells are shown. Data represent the mean and SD (bar) from three separate experiments. ( C , D ) Cells treated with BIBR at indicated concentrations or DMSO as a control for 24 h were processed to obtain cytoplasmic and nuclear extracts. Representative Western blots showing cytoplasmic and nuclear protein levels of TERT, MYC, TRF2, and α-tubulin in 4134/Late ( C ) and BL41 ( D ) cells are shown. α-tubulin and TRF2 were used as loading controls for the cytoplasmic and nuclear fractions, respectively. The original Western blots are shown in . Graphs next to the blots show the values in arbitrary units of densitometric analysis performed with ImageJ software. Data represent the mean and SD (bar) from three separate experiments. A significant difference between values in BIBR-treated vs. DMSO-treated cells is shown: * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: The expression of TERT, MYC, p65, phosphorylated p65 (p-p65), cyclin-dependent kinase inhibitor 1A (CDKN1A, P21), telomeric repeat binding factor 2 (TRF2), and α-tubulin were evaluated by the following antibodies: TERT (ab32020, Abcam, Cambridge, UK; abx120550, Abexxa, Arlington, TX, USA), NF-κB p65 (ab32536, Abcam), phospho-NF-κB p65 (Ser536) (93H1, Cell Signaling), MYC (ab32072, Abcam), P21 (ab109520, Abcam), TRF2 (NB110-57130, Novus Biological, Littleton, CO, USA), GAPDH (GTX100118, GeneTex, Alton Pkwy, Irvine, CA, USA), and α-tubulin (Sigma-Aldrich).

Techniques: Reverse Transcription, Inhibition, Control, Expressing, Western Blot, Software

Journal: Cancers

Article Title: Short-Term TERT Inhibition Impairs Cellular Proliferation via a Telomere Length-Independent Mechanism and Can Be Exploited as a Potential Anticancer Approach

doi: 10.3390/cancers15102673

Figure Lengend Snippet: p65 inhibition recapitulated the effects of TERT inhibition on MYC and the cell cycle. ( A , B ) Representative Western blots showing total protein levels of p-p65 and MYC in 4134/Late ( A ) and BL41 ( B ) cells upon treatment with Ammonium pyrrolidine dithiocarbamate (PDTC) at the indicated concentrations for 24 h. α-tubulin was used as a loading control. The original Western blots are shown in . Graphs on the right show the values in arbitrary units of densitometric analysis performed with ImageJ software. Data represent the mean and SD (bar) from two separate experiments. ( C , D ) 4134L/Late ( C ) and BL41 ( D ) cells were treated with 30 μM BIBR, PDTC at indicated concentrations, or DMSO as a control for 24 h, labelled with propidium iodide (PI) and analysed by flow cytometry for cell cycle profiles. Panels from a representative experiment are shown. Graphs on the right show percentages of cells in G1, S, and G2/M-phases, respectively. Values show the means and SD (bar) of three separate experiments. A significant difference between values in BIBR-treated or PDTC-treated cells vs. DMSO-treated cells is shown: * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The expression of TERT, MYC, p65, phosphorylated p65 (p-p65), cyclin-dependent kinase inhibitor 1A (CDKN1A, P21), telomeric repeat binding factor 2 (TRF2), and α-tubulin were evaluated by the following antibodies: TERT (ab32020, Abcam, Cambridge, UK; abx120550, Abexxa, Arlington, TX, USA), NF-κB p65 (ab32536, Abcam), phospho-NF-κB p65 (Ser536) (93H1, Cell Signaling), MYC (ab32072, Abcam), P21 (ab109520, Abcam), TRF2 (NB110-57130, Novus Biological, Littleton, CO, USA), GAPDH (GTX100118, GeneTex, Alton Pkwy, Irvine, CA, USA), and α-tubulin (Sigma-Aldrich).

Techniques: Inhibition, Western Blot, Control, Software, Flow Cytometry

Journal: Cancers

Article Title: Short-Term TERT Inhibition Impairs Cellular Proliferation via a Telomere Length-Independent Mechanism and Can Be Exploited as a Potential Anticancer Approach

doi: 10.3390/cancers15102673

Figure Lengend Snippet: BIBR treatment downregulated the expression of zebrafish myca and mycb and upregulated p21 . Twelve hours post fertilization (hpf), zebrafish wild-type (WT) and tert mutant ( tert hu3430/hu3430 ; tert −/−) embryos were treated with 2 μM BIBR or DMSO as a control for 12 h, from 12 to 24 hpf. Levels of relative mRNA expression for the indicated genes in WT ( A – C ) and tert mutant ( D – F ) embryos are shown. Data represent the mean and SD (bar) from three separate experiments. ( G ) Representative Western blots showing total protein levels of Myc in WT and tert mutant zebrafish embryos upon treatment with 2 μM BIBR for 12 h. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as a loading control. The original Western blots are shown in . The graph shows the values in arbitrary units of densitometric analysis performed with ImageJ software. Data represent the mean and SD (bar) from two separate experiments. A significant difference between values in BIBR-treated embryos vs. DMSO-treated embryos is shown: * p < 0.05; *** p < 0.001; ns: not significant.

Article Snippet: The expression of TERT, MYC, p65, phosphorylated p65 (p-p65), cyclin-dependent kinase inhibitor 1A (CDKN1A, P21), telomeric repeat binding factor 2 (TRF2), and α-tubulin were evaluated by the following antibodies: TERT (ab32020, Abcam, Cambridge, UK; abx120550, Abexxa, Arlington, TX, USA), NF-κB p65 (ab32536, Abcam), phospho-NF-κB p65 (Ser536) (93H1, Cell Signaling), MYC (ab32072, Abcam), P21 (ab109520, Abcam), TRF2 (NB110-57130, Novus Biological, Littleton, CO, USA), GAPDH (GTX100118, GeneTex, Alton Pkwy, Irvine, CA, USA), and α-tubulin (Sigma-Aldrich).

Techniques: Expressing, Mutagenesis, Control, Western Blot, Software

Journal: Frontiers in Nutrition

Article Title: Astaxanthin alleviates altered hepatic lipid metabolism and oxidative stress in animals fed a high-sucrose diet

doi: 10.3389/fnut.2026.1781406

Figure Lengend Snippet: (A) Representative photomicrographs of p-NFκB p65 immunocytochemical staining in liver tissues (400 × magnification), and (B) quantitative immunohistochemical analysis of p-NFκB p65 expression in the liver of rats fed a reference diet (RD), RD plus ASTX (RD + ASTX), high-sucrose diet (HSD), and HSD plus ASTX (HSD + ASTX). Values (mean ± SEM, n = 6 ) are expressed as the in tegrated optical density (IOD). Values that do not share the same letter were significantly different ( p < 0.05) when one variable at a time was compared by two-way ANOVA. D, effect of Diet. I, effect of intervention. Int (D x I), interaction between diet x intervention. ns, not significant.

Article Snippet: The samples were incubated in a humid chamber first with a specific primary antibody for Nrf2 (mouse monoclonal antibody; sc-365949; Santa Cruz Biotechnology) and p-NFκB p65 (mouse monoclonal antibody; sc-136548; Santa Cruz Biotechnology) (for 14–16 h at 4 °C) and then with biotin-conjugated secondary antibody (antimouse, 1:100 dilution, Sigma) for 30 min at room temperature.

Techniques: Staining, Immunohistochemical staining, Expressing

Journal: Frontiers in Nutrition

Article Title: Astaxanthin alleviates altered hepatic lipid metabolism and oxidative stress in animals fed a high-sucrose diet

doi: 10.3389/fnut.2026.1781406

Figure Lengend Snippet: Mechanisms by which ASTX modulates liver lipid metabolism and oxidative stress in HSD-fed rats. ASTX supplementation mitigates hepatic injury by reducing steatosis and triglyceride accumulation, mainly through enhanced mitochondrial fatty acid β-oxidation (↑ CPT-1 activity) without affecting lipogenesis. In parallel, ASTX improves hepatic redox homeostasis by lowering ROS levels, increasing antioxidant enzyme activities (CAT, GST) and restoring GSH content. Increased Nrf2 and decreased p-NFκB p65 protein expression were also modulated by ASTX treatment. Solid lines indicate effects supported by the experimental data, whereas dashed lines represent suggested pathways not directly evaluated in this study.

Article Snippet: The samples were incubated in a humid chamber first with a specific primary antibody for Nrf2 (mouse monoclonal antibody; sc-365949; Santa Cruz Biotechnology) and p-NFκB p65 (mouse monoclonal antibody; sc-136548; Santa Cruz Biotechnology) (for 14–16 h at 4 °C) and then with biotin-conjugated secondary antibody (antimouse, 1:100 dilution, Sigma) for 30 min at room temperature.

Techniques: Activity Assay, Expressing

Journal: Frontiers in Nutrition

Article Title: Cannabis oil modulates liver alterations and endocannabinoid system changes in a female rat model of diet-induced MASLD

doi: 10.3389/fnut.2026.1770150

Figure Lengend Snippet: Liver NrF2 and NFκB p65 expression in female rats fed a reference diet (RD), sucrose-rich diet (SRD) or SRD with cannabis oil (SRD + Ca). (A) Representative photomicrographs of immunocytochemical staining NrF2 in the liver sections of rats. Decreased levels of nuclear and cytoplasmic positive markers are observed in the SRD group. Scale bar 50 μm. (B) Quantitative immunohistochemical analysis of liver NrF2 expression expressed as integrated optical density (IOD). (C) Representative photomicrographs of immunocytochemical staining NFκB p65 in the liver sections of rats. Increased levels of nuclear and cytoplasmic positive markers are observed in the SRD group. Scale bar 50 μm. (D) Quantitative immunohistochemical analysis of NFκB p65 expression in the liver. Data are expressed as mean ± SEM ( n = 6). Statistical differences were evaluated by one-way ANOVA followed by the Newman–Keuls post hoc test (* P < 0.05).

Article Snippet: The samples were incubated in a humid chamber first with a specific primary antibody for 4-HNE (mouse monoclonal antibody; Catalog # MAB3249; R&D Systems), NrF2 (mouse monoclonal antibody; sc-365949; Santa Cruz Biotechnology), NF-κB p65 (mouse monoclonal antibody; sc-8008; Santa Cruz Biotechnology) and TGF-β1 (mouse monoclonal antibody; sc-52893; Santa Cruz Biotechnology) (for 14–16 h at 4°C) and then with biotin-conjugated secondary antibody (anti-mouse, 1:100 dilution, Sigma) for 30 min at room temperature.

Techniques: Expressing, Cannabis, Staining, Immunohistochemical staining

Journal: Scientific Reports

Article Title: T cell factor-4 functions as a co-activator to promote NF-κB-dependent MMP-15 expression in lung carcinoma cells

doi: 10.1038/srep24025

Figure Lengend Snippet: ( a ) TCF-4 potentiates the binding between NF-κB p65 and the MMP-15 promoter. The LLC cells were transfected with si-NC (20 nmol/ml), si-TCF4-1 (20 nmol/ml) or si-TCF4-2 (20 nmol/ml) for 36 hours, and then harvested for ChIP assay. The cell lysates were immunoprecipitated by p65 antibody or rabbit IgG as control. ( b ) TCF-4 interacts with NF-κB p65 which binds to the promoter of MMP-15. The LLC cells were transfected with si-NC (20 nmol/ml), si-TCF4-1 (20 nmol/ml) or si-TCF4-2 (20 nmol/ml) for 36 hours, and then harvested for ChIP assay. The cell lysates were immunoprecipitated by TCF-4 antibody or rabbit IgG as control. ( c ) Protein-protein interaction between TCF-4 and NF-κB p65 in LLC cells. The LLC cells were transfected with si-NC (20 nmol/ml), si-TCF4-1 (20 nmol/ml) or si-TCF4-2 (20 nmol/ml) for 36 hours, and then harvested for immunoprecipitation (IP) and Western blotting assay of p65 protein. The cell lysates were immunoprecipitated by TCF-4 antibody or rabbit IgG as control. ( d ) Protein-protein interaction between TCF-4 and NF-κB p65 in SAEC cells. The SAEC cells were transfected with PCMV (0.4 μg/ml) or PCMV-TCF4 (0.4 μg/ml) for 36 hours, and then collected for IP and Western blotting assay of p65. The cell lysates were immunoprecipitated by TCF-4 antibody or rabbit IgG as control. Experiment from ( a – d ) was repeated for 3 times, and the representative results were displayed.

Article Snippet: Mouse endogenous p65 was knocked down by using the two mixed commercial siRNAs (sc-29411 and sc-44213, Santa Cruz).

Techniques: Binding Assay, Transfection, Immunoprecipitation, Control, Western Blot

Journal: Scientific Reports

Article Title: T cell factor-4 functions as a co-activator to promote NF-κB-dependent MMP-15 expression in lung carcinoma cells

doi: 10.1038/srep24025

Figure Lengend Snippet: ( a ) The mRNA levels of MMP-15, TCF-4 and p65 in the LLC cells transfected with PCMV (0.4 μg/ml) or PCMV-TCF-4 (0.4 μg/ml) plus si-NC (20 nmol/ml) or a siRNA for mouse NF-κB p65 (si-p65, 20 nmol/ml) for 36 hours (n = 5, **P < 0.01). ( b ) The mRNA levels of MMP-15, TCF-4 and p65 in the LLC cells transfected with si-NC (20 nmol/ml) or si-TCF4-1 (20 nmol/ml) plus pCDNA3.1 (0.4 μg/ml) or pCDNA-p65 (0.4 μg/ml) for 36 hours (n = 5, **P < 0.01). ( c ) Immunoblotting assay of TCF-4 and p65 in the cytosol or nucleus of LLC cells transfected with si-NC (20 nmol/ml) or si-TCF4-1 (20 nmol/ml) for 36 hours. ( d ) Relative luciferase activity of LLC cells transfected with si-NC (20 nmol/ml) or si-TCF4-1 (20 nmol/ml) plus a reporter construct (0.4 μg/ml) containing the promoter of CCL20 for 36 hours (n = 4, **P < 0.01). The tests from ( a – d ) were repeated for 3 times, and the representative results were displayed.

Article Snippet: Mouse endogenous p65 was knocked down by using the two mixed commercial siRNAs (sc-29411 and sc-44213, Santa Cruz).

Techniques: Transfection, Western Blot, Luciferase, Activity Assay, Construct

Journal: Biology direct

Article Title: Hypoxia-induced activation of HIF-1alpha/IL-1beta axis in microglia promotes glioma progression via NF-κB-mediated upregulation of heparanase expression.

doi: 10.1186/s13062-024-00487-w

Figure Lengend Snippet: Fig. 4 IL-1β from hypoxic microglia promotes glioma progression and causes activation of NF-κB and upregulation of HPSE in vivo. (A and B) Changes in tumor volume and weight were shown. (C) Relative protein levels of p-p65, p65, and HPSE, as quantified using western blot. (D) The histological mor phology of tumor tissue was detected by HE staining. Relative protein levels of PCNA, E-cadherin, and vimentin, as quantified using IHC. n = 5. *P < 0.05, **P < 0.01,***P < 0.001

Article Snippet: After blocking, the membranes were treated with antibodies against HIF-1α (ab179483; Abcam, Cambridge, MA, USA), IL-1β (ab315084; Abcam, Cambridge, MA, USA), p-p65 (#3033; Cell Signaling Technology, Boston, MA, USA), p65 (#4764; Cell Signaling Technology, Boston, MA, USA), HPSE (DF12411; Affinity Biosciences, Changzhou, Jiangsu, China), and anti-β-actin (AF7018; Affinity Biosciences, Changzhou, Jiangsu, China) at 4 °C overnight.

Techniques: Activation Assay, In Vivo, Western Blot, Staining

Journal: Biology direct

Article Title: Hypoxia-induced activation of HIF-1alpha/IL-1beta axis in microglia promotes glioma progression via NF-κB-mediated upregulation of heparanase expression.

doi: 10.1186/s13062-024-00487-w

Figure Lengend Snippet: Fig. 5 IL-1β increases heparanase expression in glioma cells by activating NF-κB. (A and B) The level of p-p65/p65 in U251 and U87 cells was monitored by western blot. (A and C) The abundance of HPSE protein in U251 and U87 cells was evaluated by western blot. n = 3. ***P < 0.001

Article Snippet: After blocking, the membranes were treated with antibodies against HIF-1α (ab179483; Abcam, Cambridge, MA, USA), IL-1β (ab315084; Abcam, Cambridge, MA, USA), p-p65 (#3033; Cell Signaling Technology, Boston, MA, USA), p65 (#4764; Cell Signaling Technology, Boston, MA, USA), HPSE (DF12411; Affinity Biosciences, Changzhou, Jiangsu, China), and anti-β-actin (AF7018; Affinity Biosciences, Changzhou, Jiangsu, China) at 4 °C overnight.

Techniques: Expressing, Western Blot

Journal: Genes & Diseases

Article Title: A novel FGFR1 inhibitor CYY292 suppresses tumor progression, invasion, and metastasis of glioblastoma by inhibiting the Akt/GSK3β/snail signaling axis

doi: 10.1016/j.gendis.2023.02.035

Figure Lengend Snippet: CYY292 suppresses the stemness of GBM cells. (A, C) Immunoblot analyses of Nanog and Sox 2 levels in U87MG and LN229 cells treated with vehicle or CYY292 at 0.3, 0.5, and 1 μM for 24 h ( n = 3). (B, D) qPCR analysis of Nanog, Sox2, and Oct 4 levels in cells treated as described in (A, C) ( n = 3). (E, G) U87MG and LN229 cells were seeded in six-well plates and treated with 0.5 or 1 μM CYY292 or vehicle for colony formation assay. (F, H) Quantification of the U87MG cells and LN229 as shown in (E, G). (I, K) Immunoblot analyses of pIκBα/IκBα, p-p65/p65, and p-p38/p38 levels in U87MG and LN229 cells treated with vehicle or CYY292 at 0.3, 0.5, and 1 μM for 24 h. β-Actin was used as a protein-loading protein. (J, L) qPCR analysis of the levels of chemokine cytokines (IL1β, IL6, IL8, IL11) and chemokines (CCl2, CCl5) in cells treated as described in (A, C) ( n = 3). All data are presented as means ± SD ( n = 3 independent experiments). ∗ P < 0.05, ∗∗ P < 0.01, # P > 0.05. Differences are tested using one-way analysis of variance (ANOVA) with Tukey's post hoc test.

Article Snippet: The cell lysates were subjected to immunoblotting using anti-FGFR1 (Cell Signaling, #9740, 1:1000), anti-FGFR2 (Cell Signaling, #23328, 1:1000), anti-FGFR3 (Cell Signaling, #4574, 1:1000), anti-phosphorylated FGFR2 (Cell Signaling, #3476, 1:1000), anti-glial fibrillary acidic protein (GFAP) (Cell Signaling, #3670, 1:200), anti-AKT (Cell Signaling, #4691, 1:2000), anti-phosphorylated AKT (Cell Signaling, #4060, 1:1000), anti-p44/42 MAPK (Cell Signaling, #4695, 1:3000), anti-phosphorylated p44/42 MAPK (Cell Signaling, #4370, 1:3000), anti-GAPDH (Cell Signaling, #97166, 1:5000), anti-Vimentin (Cell Signaling, #5741, 1:3000), anti-GSK3β (Cell Signaling, #5676, 1:2000), anti-phosphorylated GSK3β (Cell Signaling, #9322, 1:2000), anti-snail (Cell Signaling, #3879, 1:1000), anti-slug (Cell Signaling, # 9585.1:1000), anti-sox 2 (Cell Signaling, #3579, 1:1000), anti-Nanog (Cell Signaling, #4903.1:1000), anti-IκBɑ (Cell Signaling, #9242.1:2000), anti-phosphorylated IκBɑ (Cell Signaling, #5209.1:2000), anti-p65 (Cell Signaling, #8242, 1:500), anti-phosphorylated p65 (Cell Signaling, #13346,1:500), anti-p38 (Cell Signaling, #8690.1:1000), anti-phosphorylated p38 (Cell Signaling, #4511, 1:1000), anti-cleaved caspase 3 (Cell Signaling, #9661.1:200), anti-β-actin (Cell Signaling, #4970.1:5000), anti-phosphorylated FGFR1 (Abcam, ab173305, 1:1000), anti-phosphorylated FGFR3 (Abcam, ab155960,1:1000), anti-CD31 (Abcam, ab28364, 1:200), anti-F4/80 (Abcam, ab11101, 1:200), anti-phosphorylated serine (Abcam, ab9332, 1:1000), anti-ki67 (Abcam, ab15580, 1:300), anti-mouse (Abcam, ab6728,1:10,000), and anti-rabbit (Abcam, ab6721, 1:10,000).

Techniques: Western Blot, Colony Assay

Journal: Dose-Response

Article Title: Ginsenoside Rh2 Suppresses the Fanconi Anemia Pathway by Inhibiting NF-κB-Mediated FANCL Transcription in Bladder Cancer

doi: 10.1177/15593258251411761

Figure Lengend Snippet: Rh2 blocks NF-κB signaling pathway in bladder cancer. (A-C) Western blot assay show the expression of FANCL as well as the phosphorylation of p65 (Ser536), β-catenin (Ser37), Akt (Thr308), JNK (Thr183/Tyr185), STAT3 (Tyr705) to determine the activity of NF-κB, Wnt/β-catenin, PI3K/Akt, JNK/AP-1 and JAK2/STAT3 pathways in T24 (A), 5637 (B), and RT4 (C) bladder cancer cell lines additionally treated with CHIR99021 (NF-κB-inducing kinase, GSK3 inhibitor), SC79 (Akt activator), anisomycin (JNK activator), colivelin (STAT3 activator). (D-F) The relative mRNA levels of FANCL are detected by qPCR in T24 (D), 5637 (E), and RT4 (F) bladder cancer cell lines additionally treated with agonists of the pathway mentioned above. “*” indicates significant difference ( P < .05) compared with “Cisplatin”, and “#” indicates significant difference ( P < .05) compared with “Cisplatin/Rh2”

Article Snippet: Membranes were incubated with the following primary antibodies at 1:2000 dilution overnight at 4°C: FANCI (Thermo Fisher Scientific, Cat. A300-212 A), FANCD2 (Thermo Fisher Scientific, Cat. MA1-16570), FANCP/SLX4 (Thermo Fisher Scientific, Cat. PA5-45039), FANCQ/XPF (Thermo Fisher Scientific, Cat. PA5-117118), PCNA (Thermo Fisher Scientific, Cat. 13-3900), FANCA (Thermo Fisher Scientific, Cat. 14657), FANCC (Thermo Fisher Scientific, Cat. H00002176-B01P), FANCM (Thermo Fisher Scientific, Cat. PA5-68191), FANCF (Abcam, USA, Cat. ab105266), FAAP24 (Abcam, Cat. ab122800), FAAP100 (Abcam, Cat. ab224178), FANCL (Abcam, Cat. ab272618), FANCB (Abcam, Cat. ab84536), USP1 (Cell Signaling Technology, USA, Cat. 4933), UAF1/WDR48 (Abcam, Cat. ab97343), GAPDH (Cell Signaling Technology, Cat. 2118), p65 (Cell Signaling Technology, Cat. 8242), p-p65 Ser536 (Cell Signaling Technology, Cat. 3033), β-catenin (Cell Signaling Technology, Cat. 8480), p-β-catenin Ser37 (Cell Signaling Technology, Cat. 9561), Akt (Cell Signaling Technology, Cat. 9272), p-Akt Thr308 (Cell Signaling Technology, Cat. 9275), JNK (Cell Signaling Technology, Cat. 9252), p-JNK Thr183/Tyr185 (Cell Signaling Technology, Cat. 4671), STAT3 (Cell Signaling Technology, Cat. 4904), and p-STAT3 Tyr705 (Cell Signaling Technology, Cat. 9145).

Techniques: Western Blot, Expressing, Phospho-proteomics, Activity Assay

Journal: Dose-Response

Article Title: Ginsenoside Rh2 Suppresses the Fanconi Anemia Pathway by Inhibiting NF-κB-Mediated FANCL Transcription in Bladder Cancer

doi: 10.1177/15593258251411761

Figure Lengend Snippet: Enlarged effect of cisplatin by Rh2 on bladder cancer in vivo. (A) Rh2 enhances cisplatin efficacy in a T24 xenograft model (n = 5), significantly reducing tumor volume. (B) Western blot assay show the mono-ubiquitination of FANCD2, FANCI, the phosphorylation of p65 as well as the expression of FANCL in xenografted tumor. Rh2 inhibits ID2 complex monoubiquitination, suppresses FANCL expression, and attenuated NF-κB signaling, as shown by decreased p-p65 levels. (C, D) Immunohistochemistry show the positive staining of FANCL (C) and p-p65 (D) in xenografted tumors. Scale bar, 0.02 mm. “*” indicates significant difference ( P < .05) compared with “NC”, and “#” indicates significant difference ( P < .05) compared with “Cisplatin”, and “^” indicates significant difference ( P < .05) compared with “Cisplatin/Rh2”

Article Snippet: Membranes were incubated with the following primary antibodies at 1:2000 dilution overnight at 4°C: FANCI (Thermo Fisher Scientific, Cat. A300-212 A), FANCD2 (Thermo Fisher Scientific, Cat. MA1-16570), FANCP/SLX4 (Thermo Fisher Scientific, Cat. PA5-45039), FANCQ/XPF (Thermo Fisher Scientific, Cat. PA5-117118), PCNA (Thermo Fisher Scientific, Cat. 13-3900), FANCA (Thermo Fisher Scientific, Cat. 14657), FANCC (Thermo Fisher Scientific, Cat. H00002176-B01P), FANCM (Thermo Fisher Scientific, Cat. PA5-68191), FANCF (Abcam, USA, Cat. ab105266), FAAP24 (Abcam, Cat. ab122800), FAAP100 (Abcam, Cat. ab224178), FANCL (Abcam, Cat. ab272618), FANCB (Abcam, Cat. ab84536), USP1 (Cell Signaling Technology, USA, Cat. 4933), UAF1/WDR48 (Abcam, Cat. ab97343), GAPDH (Cell Signaling Technology, Cat. 2118), p65 (Cell Signaling Technology, Cat. 8242), p-p65 Ser536 (Cell Signaling Technology, Cat. 3033), β-catenin (Cell Signaling Technology, Cat. 8480), p-β-catenin Ser37 (Cell Signaling Technology, Cat. 9561), Akt (Cell Signaling Technology, Cat. 9272), p-Akt Thr308 (Cell Signaling Technology, Cat. 9275), JNK (Cell Signaling Technology, Cat. 9252), p-JNK Thr183/Tyr185 (Cell Signaling Technology, Cat. 4671), STAT3 (Cell Signaling Technology, Cat. 4904), and p-STAT3 Tyr705 (Cell Signaling Technology, Cat. 9145).

Techniques: In Vivo, Western Blot, Ubiquitin Proteomics, Phospho-proteomics, Expressing, Immunohistochemistry, Staining