Journal: Frontiers in Oncology

Article Title: Proanthocyanidins-loaded complex coacervates-based drug delivery attenuates oral squamous cell carcinoma cells metastatic potential through down-regulating the Akt signaling pathway

doi: 10.3389/fonc.2022.1001126

Figure Lengend Snippet: CCDC-PA inhibited the Akt phosphorylation in HSC-3 and SCC4 cells treated with CCDC-PA at various stoichiometric ratios for 24 h. Protein lysates were subjected to Western blot analysis, and β-actin was used as the loading control (data not shown). The protein expression of Akt, JNK, MEK, Erk, and p38 as well as their phosphorylated forms were observed in (A) HSC-3 and (B) SCC4 cells. The quantification of p-Akt/Akt, p-JNK/JNK, p-MEK/MEK, p-Erk/Erk, and p-p38/p38 ratios was further done by densitometry analysis in (A) HSC-3 and (B) SCC4 cells. Control indicates a condition without treatment of CCDC-PA. 1:20, 1:10, 1:5 1:2, and 1:1 indicate the final concentration of PA in the CCDC system is equivalent to 2.5, 5.0, 10, 25, and 50 µg/mL respectively. The cropped images of blots shown in figures are used for illustrative purposes, full scan of the entire original gel(s) are included in the Supplementary Figures . Data are expressed as mean ± S.D. (n = 3), with *P< 0.05 indicating the significant difference from the controls.

Article Snippet: Blots were blocked with 5% non-fat milk at RT for 1 h, followed by overnight incubation at 4°C with specific primary antibodies at 1:1000 dilutions: ACTIN (Catalog No.: SI-A5441, Sigma); MMP2 (Catalog No.: 104577, Genetex), MMP9 (Catalog No.: 100458, Genetex), MMP12 (Catalog No.: 102928, Genetex), MMP13 (Catalog No.: 69926S, Cell Signaling), AKT (Catalog No.: 4691S, Cell Signaling), p-AKT (Catalog No.: 4060S, Cell Signaling), ERK (Catalog No.: SC-292838, Santa Cruz), p-ERK (Catalog No.: GTX129275, Genetex), JNK (Catalog No.: 474, Santa Cruz), p-JNK (Catalog No.: 9255S, Cell Signaling), MEK (Catalog No.: SC-56250, Santa Cruz), p-MEK (Catalog No.: 16500, Cell Signaling), P38 (Catalog No.: 110720, Genetex), p-P38 (Catalog No.: 133460, Genetex).

Techniques: Phospho-proteomics, Western Blot, Control, Expressing, Concentration Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Sequences of primers for each molecular gene amplification

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques:

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Differentially expression of FN1/ITGA5 in dental pulp tissue. A : GO analysis of gene expression in DPSCs from mature and young permanent dental pulp tissues. B : Immunofluorescence staining of CD31 (red) and ITGA5 (green) in mature and young permanent dental pulp tissues. C : Immunofluorescence staining of CD31 (red) and FN1 (green) in mature and young permanent dental pulp tissues. D–G: Western blot, RT-PCR, and ELISA were used to detect FN1 expression in A-DPSCs and Y-DPSCs (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Regulatory effects of FN1 in DPSCs on the biological functions of co-cultured HUVECs. A – D : After silencing FN1 in Y-DPSCs, western blot, RT-PCR, and ELISA were detected to validate FN1 expression; E – F : Cell migration of HUVECs co-cultured with DPSCs in a scratch assay. G – I : The tube formation assay was performed by counting branch points and capillary length. J : Proliferative ability was determined using CCK-8 assay (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Migration, Wound Healing Assay, Tube Formation Assay, CCK-8 Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: FN1 regulates the tube formation function of HUVECs via ITGA5. A / B : Cell migration of HUVECs treatment with FN1 or siR-ITGA5 in scratch assay. C – E : The tube formation assay of HUVECs treatment with FN1 or siR-ITGA5 was performed by counting branch points and capillary length. F : Proliferative ability was determined using CCK-8 assay (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Migration, Wound Healing Assay, Tube Formation Assay, CCK-8 Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Silencing FN1 or ITGA5 in dental pulp tissue exerts inhibitory effects on dental pulp vascular development. A : Inhibitory modulation of FN1 and ITGA5 functionality in developing dental pulp tissue was achieved through periodontal ligament injections of siRNA targeting FN1 or ITGA5, respectively, in mice. B : Immunofluorescence staining of CD31 (red) and FN1 or ITGA5 (green) in mature and young permanent dental pulp tissues. C : Numbers of vessels in dental pulp tissues of each group (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Immunofluorescence, Staining

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: FN1/ITGA5 regulates the tube formation function of HUVECs via the PI3K/AKT signaling pathway. A / B : Evaluation of PI3K/AKT signaling pathway activation status in HUVECs treated with FN1 or siRNA using Western blot. C / D : Evaluation of PI3K/AKT signaling pathway activation status in HUVECs treated with FN1 or LY294002 using Western blot. E / F : Cell migration of HUVECs treatment with FN1 or LY294002 in scratch assay. G – I : The tube formation assay of HUVECs treatment with FN1 or LY294002 was performed by counting branch points and capillary length (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Activation Assay, Western Blot, Migration, Wound Healing Assay, Tube Formation Assay

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: (A) The amino-acid sequence of the Fn14-TRAIL protein. The amino-acid sequence of the extra-cellular domain of human Fn14 (amino-acids 1-52 of the mature protein, marked in bold letters) are directly linked to the extra-cellular domain of human TRAIL (amino-acids 53-217 of the mature protein, non-bold letters). The underlined sequence represents the signal-peptide of the human Urokinase protein, utilized to secrete Fn14-TRAIL out of the cell and removed from the mature protein. (B) Fn14-TRAIL separated at denaturizing conditions on SDS-PAGE, Coomassie gel staining. (C) Western blot analysis with anti-TRAIL and anti-Fn14 primary antibodies.

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Sequencing, SDS Page, Staining, Western Blot

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: ( A ) The mRNA expression level of TRAIL, TRAIL receptors (DR4, DR5, DCR-1, DCR-2, OPG), Fn14 and TWEAK was determined by quantitive real-time PCR analysis. A representative experiment of three independent experiments is shown. Data are shown as average of triplicates (SD < 0.3), normalized against two endogenous control human genes, TBP and Actin-B, as calculated by Dataassist v2.0 software. ( B , C ) Protein expression of TRAIL, TRAIL receptors (DR4, DR5, DcR1, DcR2), Fn14 and TWEAK was determined by flow cytometeric analysis. ( D ) Fn14•TRAIL binds to HCC cells – HepG2 cells were incubated with Fn14•TRAIL, soluble TRAIL, Fn14 or the combination of the latter for 30 min at 4°c, immune-stained with PE-conjugated anti-Fn14, and analyzed by flow-cytometry. The results represent the mean +/- SD of triplicates (* p ≤ 0.05).

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Software, Incubation, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: ( A ) SK-HEP-1 [A, left panel], HepG2 [A, middle panel] and Huh7 [A, right panel] HCC cell lines, as well as NKNT3 [B, left panel] and FHB [B, right panel] hepatocyte cell lines, were incubated with 0, 3, 30 or 300 ng/ml of Fn14•TRAIL, TRAIL, Fn14-Fc or combination of the later two for 48 hours. Viable cells were stained with trypan blue and counted. The results represent the mean +/- SD of three independent experiments (* p ≤ 0.05). ( B ) HepG2 HCC cells were incubated with 30ng/ml Fn14•TRAIL for 24 hours, in the presence or absence of anti Fn14 or anti TRAIL blocking antibodies. Treated cells were stained by Annexin V-FITC and Propidium Iodide, and counted by flow cytometer (2x10 4 cells per sample). The results represent the mean +/- SD of two independent experiments (* p ≤ 0.05).

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Incubation, Staining, Blocking Assay, Flow Cytometry