Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A ) Cell viability of SKMES1 cells treated with ICG-001. ( B ) RNA level of CCL2 in SKMES1 cells. ( C ) Protein level of CCL2 in SKMES1 cells. ( D ) Cellular migration and invasion ability of THP-1 cells analyzed by transwell assay. ( E ) RNA levels of Arg-1, CD163 and IL-10 in M0 THP-1 cells treated with SKMES1 CM. ( F ) The proportion of CD206 + cells in M0 THP-1 cells with SKMES1 CM treatment. EV: empty vector, SR oe : SH3RF3 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Migration, Transwell Assay, Plasmid Preparation, Over Expression

Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A & B ) SKMES1 cells with SH3RF2 overexpression or knockdown were injected subcutaneously into the right armpit of Balb/c nude mice. Tumor size was measured every 3 days. Tumors were removed and weighed after 33 days. ( C & D ) The protein level of CCL2 in tumor determined by ELISA. ( E ) Detection of SH3RF2 expression in tumor tissues by immunohistochemistry, bar = 50 μm. ( F ) Immunohistochemistry for Ki-67 in tumor tissues, bar = 50 μm. ( G ) The proportion of CD11b + F4/80 + CD206 + cells in tumor. NC sh : negative control shRNA, SR sh−1 : SH3RF2 shRNA-1, SR sh−2 : SH3RF2 shRNA-2, EV: empty vector, SR oe : SH3RF3 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Over Expression, Knockdown, Injection, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Negative Control, shRNA, Plasmid Preparation

Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A ) Protein level of LZTS2 in SKMES1 cells with SH3RF2 and LZTS2 overexpression. ( B ) Cell viability of SKMES1 cells with SH3RF2 and LZTS2 overexpression. ( C ) Immunofluorescence detection of the distribution of β-catenin in SKMES1 cells and quantitative analysis of nuclear-to-cytoplasmic ratio of β-catenin fluorescence intensity. ( D ) Protein level of CCL2 in SKMES1 cells. ( E ) RNA levels of Arg-1, CD163 and IL-10 in M0 THP-1 cells with SKMES1 CM treatment. ( F ) Cellular migration and invasion ability of M0 THP-1 cells with SKMES1 CM treatment. ( G ) The proportion of CD206 + cells in the M0 THP-1 cells treated with SKMES1 CM. EV: empty vector, SR oe : SH3RF3 overexpression. LZ oe : LZTS2 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Over Expression, Immunofluorescence, Fluorescence, Migration, Plasmid Preparation

Journal: Journal of Nanobiotechnology

Article Title: Therapies with CCL25 require controlled release via microparticles to avoid strong inflammatory reactions

doi: 10.1186/s12951-021-00830-7

Figure Lengend Snippet: Characteristic features of PLGA particles. PLGA particles were analyzed morphologically with SEM, evaluated for endotoxin contamination with the Limulus amebocyte lysate (LAL) test, and functionally assessed by their CCL25 release profile. Representative SEM images of the PLGA particles generated are shown in a i,ii with × 40 and 200 magnification at day one, iii, iv with × 40/ × 200 magnification after 14 days of degradation, v, vi with × 40/ × 200 magnification after 28 days of degradation and vii, viii with × 1000 magnification after 63 days of degradation. Scale bars represent 500 and 100 µm, respectively. b The results of the LAL test for the detection of endotoxins are shown for different stock solutions of CCL25 (10 and 500 nM) and the supernatant from PLGA particles (CL supernatant) compared to the test standards (0.1–10 nM). c The cumulative release in percent ± SD of CCL25 from PLGA particles over time until day 63 ( n = 3) was determined by ELISA. CL CCL25-loaded, PLGA Poly (lactic-co-glycolic acid)

Article Snippet: The amount of CCL25 released from the PLGA microparticles was measured using the human CCL25/TECK DuoSet® Enzyme linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, USA).

Techniques: Generated, Enzyme-linked Immunosorbent Assay

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A serum concentration of CCL25 measured by ELISA showed a positive correlation with BMI (Pearson r 2 0.500, p<0.001). B measurement of protein levels of CCL25 measured by immunohistochemical staining showed an increase in CCL25 levels in obesity (* p<0.05 by one-way ANOVA). C Gene expression of CCL25 in adipose and liver tissue measured by rt-PCR showed greater expression in adipose tissue. D Protein levels of CCL25 were comparable between adipose and liver tissue.

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A serum concentration of CCL25 measured by ELISA in lean and obese individuals **** p<0.001 by Mann Whitney test B lymphocyte trafficking in response to pre-treatment of hepatic endothelial cells with recombinant CCL25 ****p<0.001 by two-way ANOVA

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Recombinant

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A adherence of leukocytes to HSEC after pre-treatment with CCL25 (* one-way ANOVA p<0.05) B Total CCR9 + cells isolated from normal and NAFLD liver tissue (** student’s t-test p<0.01). C CCR9 expression on leukocyte subsets from normal and NAFLD liver tissue D CCR9 expression on liver-infiltrating monocytes in normal and NAFLD liver tissue

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Isolation, Expressing

Journal: Oncotarget

Article Title: CCR9/CCL25 expression in non-small cell lung cancer correlates with aggressive disease and mediates key steps of metastasis

doi:

Figure Lengend Snippet: Serum CCL25 levels in healthy donors (n=9), SCC (n=17) and AC (n=14) patients were analyzed by ELISA. As used, the ELISA could detect >5 pg/mL of CCL25. Box plots for each group show the minimum and maximum values. The lines in the box plots indicate the median serum CCL25 concentrations of each group. The difference in CCL25 concentrations between healthy controls and SCC or AC and SCC and AC were analyzed using Mann Whitney U test and were highly significant (p < 0.0001).

Article Snippet: Serum CCL25 levels were quantified by human CCL25 Quantikine ELISA kit (R&D Systems) according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Oncotarget

Article Title: CCR9/CCL25 expression in non-small cell lung cancer correlates with aggressive disease and mediates key steps of metastasis

doi:

Figure Lengend Snippet: SCC (NCI-H520) and AC (NCI-H2126) cells were tested for their ability to migrate [A] and invade [B] toward chemotactic gradients of 0 (open bar) or 100 ng/ml (black bar) of CCL25. The cells were pre-treated with anti-human CCR9 antibody (1 μg/ml) (grey bar) during migration and invasion assays. Asterisks indicate significant differences in migration and invasion between untreated and CCL25-treated or anti-CCR9-treated cell lines (** p < 0.01). Data was analyzed using non-parametric two tailed t-test and is presented as mean +/− S.D., n=3.

Article Snippet: Serum CCL25 levels were quantified by human CCL25 Quantikine ELISA kit (R&D Systems) according to the manufacturer’s protocol.

Techniques: Migration, Two Tailed Test

Journal: Oncotarget

Article Title: CCR9/CCL25 expression in non-small cell lung cancer correlates with aggressive disease and mediates key steps of metastasis

doi:

Figure Lengend Snippet: Cells were tested for their capacity to express mRNA and protein for MMP-2 and -9 and TIMP-1 and -2. [A] SCC (NCI-H520) and AC (NCI-H2126) cells were treated for 30 min with 0 or 100 ng/mL of CCL25. Total RNA was isolated, and quantitative real time-PCR analysis was performed for mRNA expression of MMP-2 and -9 and TIMP-1 and -2. Transcript copies were presented relative to copies of 18S rRNA. [B] Active gelatinases (MMP-2 and -9) in culture supernatants were quantified by gelatin zymography. Cells were stimulated with CCL25 (0 or 100 ng/ml) for 24 h. Top: Graph represents densitometric analysis of zymography for control and treated samples, presented as band area, analyzed by ImageJ software. Bottom: Representative zymography. [C] TIMP-1 and TIMP-2 in culture supernatants were quantified by ELISA. Bars represent the concentration (pg/ml) of TIMP-1 and TIMP-2 in culture supernatants collected from cells treated with 0 or 100 ng/ml of CCL25 for 24 h. Asterisks show significant differences between untreated and CCL25-treated LuCa cells. Data was analyzed by non-parametric two tailed t-test and presented as mean +/− S.D., n=2. * p < 0.05.

Article Snippet: Serum CCL25 levels were quantified by human CCL25 Quantikine ELISA kit (R&D Systems) according to the manufacturer’s protocol.

Techniques: Isolation, Real-time Polymerase Chain Reaction, Expressing, Zymography, Control, Software, Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test