Journal: Scientific Reports

Article Title: TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells

doi: 10.1038/srep23775

Figure Lengend Snippet: All pairs of F/R primers used for real-time qPCR.

Article Snippet: Rabbit polyclonal antibodies against MMP9, SNAI1, p16 and Vimentin were bought from BioSynthesis (Beijing, China), whilst other rabbit antibodies against α-Catenin, CDK2, CDK6, Cyclin D1, SNAI2, FN1, Rb1, p53 and p21 were purchased from BosTer (Wuhan, China).

Techniques:

Journal: Journal of Orthopaedic Translation

Article Title: Pharmacological network analysis of the functions and mechanism of kaempferol from Du Zhong in intervertebral disc degeneration (IDD)

doi: 10.1016/j.jot.2023.01.002

Figure Lengend Snippet: The compound-target-signaling networks generated using Cytoscape_v3.8.0. The active compounds cyclopamine, quercetin, kaempferol and mairin could target 12 IDD disease genes including HMGB1, PTEN, IGFBP6, FGFR3, COL3A1, MMP2, SMAD2, HSPG2, GPC1, FN1, CUL4B, CSGALNACT1 (Among 13 target genes, INTS8 did not have pathway enrichment). The compound-targeted disease genes were enriched in 23 pathways.

Article Snippet: The following primary antibodies were used in this study: p16 (CSB-PA003618, CUSABIO, Wuhan, China), p21 (ab109520, Abcam, Cambridge, UK), Rb (ab224426, Abcam), hTERT (DF7129, Affinity Biotech), Nrf2 (CSB-RA225569A0HU,Cusabio), HO-1 (10701-1-AP, Proteintech), NQO-1 (11451-1-AP, Proteintech), SOD1 (10269-1-AP, Proteintech), SOD2 (ab227091, Abcam), Cleaved-caspase 3 (ab2302, Abcam), Caspase 3 (ab13847, Abcam), Bax (50599-2-Ig, Proteintech), Bcl-2 (12789-1-AP, Proteintech), aggrecan (13880-1-AP, Proteintech), collagen II (CSB-PA005739ESR2HU, Cusabio), SOX9 (ab185966, Abcam), FN1 (AF5335, Affinity Biotech), MMP2 (CSB-PA003258, Cusabio), MMP3 (17873-1-AP, Proteintech), MMP13 (CSB-PA07029A0Rb, Cusabio), ADAMTS-4 (DF6986, Affinity Biotech), ADAMTS-5 (DF13268, Affinity Biotech), p-p38 (AF4001, Affinity Biotech), p38 (14064-1-AP, Proteintech), p-JNK (AF3318, Affinity Biotech), JNK (AF6319, Affinity Biotech), p-ERK1/2 (sc-81492, Santa Cruz, Dallas, DX, USA), ERK1/2 (16443-1-AP, Proteintech).

Techniques: Generated

Journal: Journal of Orthopaedic Translation

Article Title: Pharmacological network analysis of the functions and mechanism of kaempferol from Du Zhong in intervertebral disc degeneration (IDD)

doi: 10.1016/j.jot.2023.01.002

Figure Lengend Snippet: Compound-target-signaling networks.

Article Snippet: The following primary antibodies were used in this study: p16 (CSB-PA003618, CUSABIO, Wuhan, China), p21 (ab109520, Abcam, Cambridge, UK), Rb (ab224426, Abcam), hTERT (DF7129, Affinity Biotech), Nrf2 (CSB-RA225569A0HU,Cusabio), HO-1 (10701-1-AP, Proteintech), NQO-1 (11451-1-AP, Proteintech), SOD1 (10269-1-AP, Proteintech), SOD2 (ab227091, Abcam), Cleaved-caspase 3 (ab2302, Abcam), Caspase 3 (ab13847, Abcam), Bax (50599-2-Ig, Proteintech), Bcl-2 (12789-1-AP, Proteintech), aggrecan (13880-1-AP, Proteintech), collagen II (CSB-PA005739ESR2HU, Cusabio), SOX9 (ab185966, Abcam), FN1 (AF5335, Affinity Biotech), MMP2 (CSB-PA003258, Cusabio), MMP3 (17873-1-AP, Proteintech), MMP13 (CSB-PA07029A0Rb, Cusabio), ADAMTS-4 (DF6986, Affinity Biotech), ADAMTS-5 (DF13268, Affinity Biotech), p-p38 (AF4001, Affinity Biotech), p38 (14064-1-AP, Proteintech), p-JNK (AF3318, Affinity Biotech), JNK (AF6319, Affinity Biotech), p-ERK1/2 (sc-81492, Santa Cruz, Dallas, DX, USA), ERK1/2 (16443-1-AP, Proteintech).

Techniques: Activity Assay, Migration

Journal: Journal of Orthopaedic Translation

Article Title: Pharmacological network analysis of the functions and mechanism of kaempferol from Du Zhong in intervertebral disc degeneration (IDD)

doi: 10.1016/j.jot.2023.01.002

Figure Lengend Snippet: Effects of Kaempferol IL-1β-induced ECM deposition NPCs were exposed to IL-1β stimulation (10 ​ng/ml, 24 ​h), treated with 10 ​μM kaempferol, and examined for the mRNA and protein levels of aggrecan, collagen II, SOX9, FN1 (A–B), MMP3, MMP13, ADAMTS-4, and ADAMTS-5 (C–D) using qRT-PCR and Immunoblotting, respectively. . One-way ANOVA followed by Tukey's post hoc test. N ​= ​3, ∗∗p ​< ​0.01, compared to PBS group; #p ​< ​0.05, ##p ​< ​0.01, compared with IL-1β group.

Article Snippet: The following primary antibodies were used in this study: p16 (CSB-PA003618, CUSABIO, Wuhan, China), p21 (ab109520, Abcam, Cambridge, UK), Rb (ab224426, Abcam), hTERT (DF7129, Affinity Biotech), Nrf2 (CSB-RA225569A0HU,Cusabio), HO-1 (10701-1-AP, Proteintech), NQO-1 (11451-1-AP, Proteintech), SOD1 (10269-1-AP, Proteintech), SOD2 (ab227091, Abcam), Cleaved-caspase 3 (ab2302, Abcam), Caspase 3 (ab13847, Abcam), Bax (50599-2-Ig, Proteintech), Bcl-2 (12789-1-AP, Proteintech), aggrecan (13880-1-AP, Proteintech), collagen II (CSB-PA005739ESR2HU, Cusabio), SOX9 (ab185966, Abcam), FN1 (AF5335, Affinity Biotech), MMP2 (CSB-PA003258, Cusabio), MMP3 (17873-1-AP, Proteintech), MMP13 (CSB-PA07029A0Rb, Cusabio), ADAMTS-4 (DF6986, Affinity Biotech), ADAMTS-5 (DF13268, Affinity Biotech), p-p38 (AF4001, Affinity Biotech), p38 (14064-1-AP, Proteintech), p-JNK (AF3318, Affinity Biotech), JNK (AF6319, Affinity Biotech), p-ERK1/2 (sc-81492, Santa Cruz, Dallas, DX, USA), ERK1/2 (16443-1-AP, Proteintech).

Techniques: Quantitative RT-PCR, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: Pharmacological network analysis of the functions and mechanism of kaempferol from Du Zhong in intervertebral disc degeneration (IDD)

doi: 10.1016/j.jot.2023.01.002

Figure Lengend Snippet: Table S1 The primers for qRT-PCR

Article Snippet: The following primary antibodies were used in this study: p16 (CSB-PA003618, CUSABIO, Wuhan, China), p21 (ab109520, Abcam, Cambridge, UK), Rb (ab224426, Abcam), hTERT (DF7129, Affinity Biotech), Nrf2 (CSB-RA225569A0HU,Cusabio), HO-1 (10701-1-AP, Proteintech), NQO-1 (11451-1-AP, Proteintech), SOD1 (10269-1-AP, Proteintech), SOD2 (ab227091, Abcam), Cleaved-caspase 3 (ab2302, Abcam), Caspase 3 (ab13847, Abcam), Bax (50599-2-Ig, Proteintech), Bcl-2 (12789-1-AP, Proteintech), aggrecan (13880-1-AP, Proteintech), collagen II (CSB-PA005739ESR2HU, Cusabio), SOX9 (ab185966, Abcam), FN1 (AF5335, Affinity Biotech), MMP2 (CSB-PA003258, Cusabio), MMP3 (17873-1-AP, Proteintech), MMP13 (CSB-PA07029A0Rb, Cusabio), ADAMTS-4 (DF6986, Affinity Biotech), ADAMTS-5 (DF13268, Affinity Biotech), p-p38 (AF4001, Affinity Biotech), p38 (14064-1-AP, Proteintech), p-JNK (AF3318, Affinity Biotech), JNK (AF6319, Affinity Biotech), p-ERK1/2 (sc-81492, Santa Cruz, Dallas, DX, USA), ERK1/2 (16443-1-AP, Proteintech).

Techniques:

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: Establishment and characterization of GC (Gemcitabine and Cisplatin)-resistant bladder cancer cell lines and identification of resistance-related proteins. ( A ) GC-resistant T24-R and UC3-R cell lines were generated by gradually increasing GC concentrations. Created in BioRender ( https://BioRender.com ). ( B ) Dose-response curves and calculated half-maximal inhibitory concentration (IC50) values for cisplatin (upper panel) and gemcitabine (lower panel) in parental (T24, UC3) and GC-resistant (T24-R, UC3-R) cell lines. Data are presented as the mean ± SD from at least three independent experiments. ( C ) Apoptosis rates of parental and resistant cell lines after treatment with cisplatin, as determined by flow cytometry. Data are presented as the mean ± SD ( n ≥ 3). ( D ) Transcriptomic and proteomic analyses identified FN1 (Fibronectin), EEF1A2 (Eukaryotic Translation Elongation Factor 1 Alpha 2), MRC2 (Mannose Receptor C-Type 2), RAB6B (RAB6B, Member RAS Oncogene Family), THBS1 (Thrombospondin 1), DYSF (Dysferlin), TMOD1 (Tropomodulin 1), NES (Nestin), and APOE (Apolipoprotein E) as overexpressed in GC-resistant cells. ( E ) RT-qPCR (Reverse Transcription Quantitative Polymerase Chain Reaction) and Western blot confirmed FN1 overexpression in T24-R and UC3-R. ( F ) FN1 staining was stronger in GC-resistant bladder cancer tissues (Chemotherapy Sensitive Group: n = 6; Chemotherapy Resistant Group: n = 6). For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, ** p < 0.01

Article Snippet: Following blocking with 10% normal goat serum, tissue sections were probed with a primary antibody against FN1 (HUABIO, ET1702-25), diluted at 1:200, in a 4°C overnight incubation.

Techniques: Generated, Concentration Assay, Flow Cytometry, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Staining

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: ITGB4 is critical for FN1-mediated chemotherapy resistance in bladder cancer cells. ( A ) Differential expression analysis revealed that ITGB4 (highlighted by the red box) was significantly overexpressed in T24-R cells compared to T24, suggesting a role in FN1-mediated resistance. ( B ) Structural modeling shows multiple binding sites between FN1 (blue ribbon) and ITGB4 (green ribbon). ( C ) The interaction between FN1 and ITGB4 had a binding score of −318.75 with a confidence score of 96%, indicating a stable interaction. ( D ) The Co-IP experiment confirmed that there is a mutual binding interaction between FN1 and ITGB4. ( E ) Adding rFN1 to resistant strains increased FAK (Y397) phosphorylation and inhibited apoptosis, whereas ITGB4 silencing reversed these effects, highlighting the dependency of FN1-mediated resistance on ITGB4 expression and activation. For panels ( A , C , E ), a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups

Article Snippet: Following blocking with 10% normal goat serum, tissue sections were probed with a primary antibody against FN1 (HUABIO, ET1702-25), diluted at 1:200, in a 4°C overnight incubation.

Techniques: Quantitative Proteomics, Binding Assay, Co-Immunoprecipitation Assay, Phospho-proteomics, Expressing, Activation Assay

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 silencing enhances cisplatin-induced apoptosis in bladder cancer cells. ( A ) FN1 knockdown efficiency in T24-R and UC3-R cells was confirmed by Western blot analysis. ( B ) FN1 knockdown efficiency in T24-R and UC3-R cells was confirmed by RT-qPCR analysis. ( C ) Silencing FN1 significantly increased apoptosis in the resistant cell lines by TUNEL staining. ( D ) Silencing FN1 reduced the IC50 of cisplatin in the resistant cell lines by CCK-8 assay. ( E ) In resistant cells, FN1 knockdown induced the expression of pro-apoptotic mediators, including BAX and cleaved-caspase-3, but suppressed levels of the anti-apoptotic protein Bcl-2. ( F ) Cell apoptosis rates were quantified by flow cytometry. For all panels, a t-test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, ** p < 0.01

Article Snippet: Following blocking with 10% normal goat serum, tissue sections were probed with a primary antibody against FN1 (HUABIO, ET1702-25), diluted at 1:200, in a 4°C overnight incubation.

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, TUNEL Assay, Staining, CCK-8 Assay, Expressing, Flow Cytometry

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 silencing inhibits tumor growth in vivo , enhancing cisplatin sensitivity. ( A ) Representative images of tumors from each treatment group. ( B ) Tumor volume growth curves over time measured in the T24-R xenograft model. ( C ) Apoptosis in tumor tissues was detected by TUNEL staining. ( D ) Immunohistochemical analysis showed decreased FN1 expression in tumor sections from the FN1 knockdown group. For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. * p < 0.05, *** p < 0.001

Article Snippet: Following blocking with 10% normal goat serum, tissue sections were probed with a primary antibody against FN1 (HUABIO, ET1702-25), diluted at 1:200, in a 4°C overnight incubation.

Techniques: In Vivo, TUNEL Assay, Staining, Immunohistochemical staining, Expressing, Knockdown

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 regulates FAK (Y397) phosphorylation and mediates cisplatin resistance in bladder cancer cells. ( A ) Silencing FN1 in resistant cell lines reduced the phosphorylation of FAK (Y397) as detected by Western blot. ( B ) T24 and UC3 parental and resistant cells were treated with a fixed dose of cisplatin along with a gradient of rFN1 for 48 h. Phosphorylation of FAK (Y397) was assessed by Western blot. Resistant cells (T24-R, UC3-R) showed sensitivity to rFN1 at lower concentrations under cisplatin stress. ( C ) Silencing FN1 in resistant cell lines reduced the phosphorylation of FAK (Y397) as detected by immunofluorescence. ( D ) rFN1 addition increased resistance index in resistant strains. ( E ) The addition of rFN1 reduced apoptosis in resistant strains. For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, *** p < 0.001

Article Snippet: Following blocking with 10% normal goat serum, tissue sections were probed with a primary antibody against FN1 (HUABIO, ET1702-25), diluted at 1:200, in a 4°C overnight incubation.

Techniques: Phospho-proteomics, Western Blot, Immunofluorescence