Journal: Journal of nanobiotechnology

Article Title: Evaluating the pro-survival potential of apoptotic bodies derived from 2D- and 3D- cultured adipose stem cells in ischaemic flaps.

doi: 10.1186/s12951-024-02533-1

Figure Lengend Snippet: Fig. 6 3D-ABs suppress oxidative stress and cell death, and stimulated angiogenesis in ischaemic flaps. (A) F-CHP staining for damaged collagen detec tion in the skin on POD7. Scale bars, 100 μm. (B) The three groups’ quantified F-CHP intensity (n = 6). (C) CD31 and α-SMA IF staining in the flap on POD7. Scale bars, 50 μm. (D) Quantified CD31/α-SMA-positive blood vessel density among the three groups (n = 6). (E) Dead cells in flap tissue sections on POD7 under the detection of TUNEL staining. Scale bar, 20 μm. (F) TUNEL-positive cell percentage in the dermal layer, quantified for each of the three groups (n = 6). (G) DHE staining of the skin tissues on POD7 among the three groups. Scale bars, 50 μm. (H) Quantified DHE among the three groups (n = 6). SEM error bars are used. Significance (*): p value < 0.05; equal variances ANOVA with LSD post hoc analysis or unequal variances Dunnett’s T3 technique

Article Snippet: The primary antibodies utilized were specific to CD31 (1:200), CD68 (1:200), α-SMA (1:200; Proteintech, 67735-1-Ig), iNOS (1:200), and Arg1 (1:200).

Techniques: Staining, TUNEL Assay

Journal: American Journal of Translational Research

Article Title: RNF128 suppresses the malignancy of colorectal cancer cells via inhibition of Wnt/β-catenin signaling

doi:

Figure Lengend Snippet: Loss of RNF128 increases β-catenin protein level and its target genes in colorectal cancer. (A) The mRNA expressions of cyclin D1, cyclin E1, cyclin A1 and cyclin B1 were quantified in RNF128 knockdown or overexpression cells compared to their respective controls in DLD1. (B) β-catenin, CCND1, and RNF128 expressions were confirmed using Western blotting. β-actin was used as a loading control. (C) Expression of β-catenin and RNF128 in nuclear and cytoplasmic fractions in RNF128-knockdown cells and control cells of DLD1. β-tubulin was used as the cytoplasmic reference protein. LaminB1 was used as the nuclear reference protein. *P<0.05, **P<0.01 and ***P<0.001. No significant difference (P>0.05, ns). Data represented as mean ± SD.

Article Snippet: Supernatants were collected for measuring protein concentration by BCA assay (Thermofisher Scientific, USA). β-tubulin Rabbit mAb, Lamin B1 Rabbit mAb and β-actin Rabbit mAb were purchased from Proteintech (china); β-Catenin Rabbit mAb and CCND1 Rabbit mAb were from Cell Signaling Technology (USA); Ub and RNF128 Rat mAb were from Santa Cruz Biotechnology (USA); goat anti-rabbit and goat anti-rat were from Cell Signaling Technology (USA).

Techniques: Over Expression, Western Blot, Expressing

Journal: Neoplasia (New York, N.Y.)

Article Title: Inhibition of fibronectin accumulation suppresses TUMOR growth

doi: 10.1016/j.neo.2021.06.012

Figure Lengend Snippet: Changes in fibronectin affect blood vessels and proliferation. (A) Tumor lesions exhibit less fibronectin staining in sections from knockdown (Kd) tumors compared to controls (CT). Bars represent 100 μm. (B) Kd tumors contain less fibronectin as determined by ELISA. Pieces obtained from tumor samples were weighed and lysed in protein lysis buffer to evaluate by ELISA. The amount of fibronectin was corrected to total protein measured by the BCA method. N=3/6 replicates. (C) Expression of fibronectin mRNA originating from the cancer cells (human) and the host (murine) was diminished. Using primers specific for human and murine fibronectin it was possible to differentiate between both in qPCR analyses. Human fibronectin was corrected to human HPRT and murine fibronectin to murine β-actin. N=5/3 and 7/5 for the two graphs (left to right). (D) The area of CD31 + blood vessels diminishes as does the number of vessels stained with both CD31 (in red) and the pericyte markers α-smooth muscle actin (α-SMA) or desmin (in green). Examples are shown. Bars represent 100 μm. N=11/10 for CD31 + area and stainings with αSMA and 7/7 for stainings with desmin. (E) Endothelial cells from knockdown tumors produce less fibronectin. Isolated endothelial cells were cultured in the presence of fibronectin-depleted FCS and fibronectin evaluated in the conditioned media by ELISA. N=11/5. (F) Proliferation in tumor sections was diminished in Kd tumors as evidenced by the decrease in the percentage of Ki67 + cells. N=9/9. Examples are shown. Bars represent 100 μm. (G) Tumor sections were stained using TUNEL to mark apoptotic cells. No difference was seen between CT and Kd tumors. N=3/3. Examples are shown. Bars represent 100 μm. All pairs were evaluated using t-test. * P <0.05, ** P <0.005, *** P <0.0001.

Article Snippet: We also used rat anti-mouse CD31 (BIO-RAD; MCA2388GA; 1:100), mouse anti-alpha-smooth muscle actin (α-SMA)-Cy3 (Merck; #A2547; 1:200), rabbit anti-desmin (Dianova; #DLN-13732; 1:100), rabbit monoclonal antibody against YAP (Cell signaling #14074, 1:200), goat anti-rabbit-Alexa 647 (Abcam #ab150079; 1:1000), goat anti-mouse-Cy2 (Dianova; #115-225-166; 1:1000), goat anti-rat-Alexa 594 (Dianova; #112-585-062; 1:1000).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Lysis, Expressing, Isolation, Cell Culture, TUNEL Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: Inhibition of fibronectin accumulation suppresses TUMOR growth

doi: 10.1016/j.neo.2021.06.012

Figure Lengend Snippet: Histology of pUR4-treated tumors confirms changes in matrix and reveals a decrease in proliferation. (A) Stained tumor sections show a decrease in fibronectin and collagen in pUR4-treated animals and no changes in R1R2-treated animals. Bars represent 100 μm. (B) Western blots from the tumors confirm a decrease in fibronectin and collagen with pUR4 treatment, but collagen I did not decrease with R1R2 therapy. N=4/4/4/4 replicates/treatment. Relevant pairs were evaluated by t-tests. * P <0.05. (C) Proliferation was diminished after treatment with pUR4. Sections were stained for Ki67. Bars represent 100 μm. N=9/9/9/9. Pairs were evaluated by t-tests. ** P <0.01. (D) Apoptosis was not affected in vivo by the various treatments. Sections were stained for TUNEL. Examples are shown. Bars represent 100 μm. N=9/9/9/9. Pairs were evaluated by t-tests. (E) Evaluation of blood vessels shows a decrease in area of CD31 + vessels with pUR4 treatment as well as the number of CD31 + αSMA + or CD31 + desmin + double positive vessels (adjusted to the area). Sections from representative tumors with bars representing 100μm. These are intratibial tumors from mice treated for 10 days with the peptides. N=12/12/11/12 for CD31 alone or with α-smooth muscle actin (αSMA) and 11/10/7/7 for staining with desmin. Pairs were compared by t-test. * P <0.05, **** P <0.0001.

Article Snippet: We also used rat anti-mouse CD31 (BIO-RAD; MCA2388GA; 1:100), mouse anti-alpha-smooth muscle actin (α-SMA)-Cy3 (Merck; #A2547; 1:200), rabbit anti-desmin (Dianova; #DLN-13732; 1:100), rabbit monoclonal antibody against YAP (Cell signaling #14074, 1:200), goat anti-rabbit-Alexa 647 (Abcam #ab150079; 1:1000), goat anti-mouse-Cy2 (Dianova; #115-225-166; 1:1000), goat anti-rat-Alexa 594 (Dianova; #112-585-062; 1:1000).

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