Journal: Cell Biology and Toxicology

Article Title: SENP3 mediated DeSUMOylation of macrophage derived CCL17 accelerates atherosclerosis via regulation of Treg

doi: 10.1007/s10565-025-10099-3

Figure Lengend Snippet: Macrophages stimulated by oxLDL affect the chemotaxis of Treg through CCL17 and CCL22 competition. Macrophages were stimulated with oxLDL, and Treg cells were co-cultured with macrophage supernatant. A Immunofluorescence detection of co-localization of CCL22 and macrophages (CD68) in plaque tissue of AS mice. B The content of CCL22 in the supernatant of macrophages was detected by ELISA. C Transwell detected the effect of macrophages on the chemotaxis of Treg. ( D - E ) Transwell detected the migration of Treg cells. * p < 0.05, ** p < 0.01, *** p < 0.001. N = 3

Article Snippet: Prior to the transwell migration assay, the induced Tregs were treated with recombinant mouse CCL17 (MCE, HY-P71891A) or recombinant mouse CCL22 (MCE, HY-P7248) or anti-CCL17 (R&D, Catalog #: MAB529) and anti-CCL22 antibodies (R&D, Catalog #: MAB529).

Techniques: Chemotaxis Assay, Cell Culture, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Migration

Journal: The Journal of Clinical Investigation

Article Title: TRAIL agonists rescue mice from radiation-induced lung, skin, or esophageal injury

doi: 10.1172/JCI173649

Figure Lengend Snippet: ( A ) Experimental timeline. Female C57BL/6 mice orthotopically injected with e0771 on day 0 and when tumors reached 2–5 mm in size mice were irradiated with a whole-thorax irradiation dose of 20 Gy and treated with either TLY012, ONC201, or the combination ( n = 3–4 per treatment per group). (Created in BioRender. Strandberg J. 2024. https://BioRender.com/c65v064. ) ( B and C ) Tumors were removed after mice were euthanized 18 days after cell injection, and weight and volume were calculated. ( D ) Pulse oximetry readings before radiation and 9 days after radiation showed oxygen saturation more conserved in the TLY012-treated group compared with irradiated controls. Values are mean ± SEM. ( E ) Statistical analysis of cytokine fold change showed significant decrease in levels of MDC/CCL22 ( P = 0.035) in comparison with irradiated controls (1-way ANOVA with Tukey’s post hoc test).

Article Snippet: Seven-week-old Trail –/– female mice received a single thoracic x-ray irradiation dose of 20 Gy with shielding of other organs and were treated with either 20 μg of anti-CCL22 antibody (AF439, R&D Systems Inc.) dissolved in 500 μL of sterile normal saline via i.p. injection every other day for 2 weeks or 20 μg of polyclonal goat IgG (AB-108-C, R&D Systems Inc.) dissolved in 500 μL of normal saline via i.p. injection every other day for 2 weeks ( n = 5 per treatment per group).

Techniques: Injection, Irradiation, Comparison

Journal: The Journal of Clinical Investigation

Article Title: TRAIL agonists rescue mice from radiation-induced lung, skin, or esophageal injury

doi: 10.1172/JCI173649

Figure Lengend Snippet: ( A ) Experimental timeline of Trail –/– female mice treated with 20 μg of anti-CCL22 in 500 μL of saline or 20 μg of goat IgG in 500 μL of saline every other day for 2 weeks ( n = 5 per treatment per group). (Created in BioRender. Strandberg J. 2024. https://BioRender.com/j52q830. ) ( B ) H&E images of each mouse 2 weeks after thoracic irradiation (original magnification, ×20). Scale bar: 100 μm. ( C ) Quantification of inflammatory scores provided by a blinded pathologist showed decrease in inflammation in Trail –/– female mice treated with anti-CCL22 ( n = 5 per treatment per group) but not to a significant extent (1-tailed Mann-Whitney test) ( P = 0.1032). Values are mean ± SEM.

Article Snippet: Seven-week-old Trail –/– female mice received a single thoracic x-ray irradiation dose of 20 Gy with shielding of other organs and were treated with either 20 μg of anti-CCL22 antibody (AF439, R&D Systems Inc.) dissolved in 500 μL of sterile normal saline via i.p. injection every other day for 2 weeks or 20 μg of polyclonal goat IgG (AB-108-C, R&D Systems Inc.) dissolved in 500 μL of normal saline via i.p. injection every other day for 2 weeks ( n = 5 per treatment per group).

Techniques: Saline, Irradiation, MANN-WHITNEY

Journal: Chemical Engineering Journal

Article Title: Microfluidic fabrication of lipid nanoparticles for co-delivery of siRNA and hydroxychloroquine: An engineered theranostic platform for enhanced breast cancer treatment

doi: 10.1016/j.cej.2025.160172

Figure Lengend Snippet: Fig. 3. Effect of various treatments on cell proliferation. Screening of human- (A) and mouse- (B) derived candidate siRNA sequences. (C) Cell counting kit-8 (CCK-8) assay to evaluate anti-proliferative results against 4T1 cells. (D) Crystalline violet staining to visualize agent-treated cell colonies. (E) Ultraviolet absorption of crystalline violet eluents from stained colonies (n = 3). *p<0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The LysoTracker Green and Autophagy Staining Assay Kit with MDC were obtained from Beyotime (Shanghai, China).

Techniques: Derivative Assay, Cell Counting, CCK-8 Assay, Staining

Journal: PLoS ONE

Article Title: LncRNA HOTAIR promotes proliferation, invasion and migration in NSCLC cells via the CCL22 signaling pathway

doi: 10.1371/journal.pone.0263997

Figure Lengend Snippet: A, B: the expression of HOTAIR and CCL22 mRNA were detected by RT-qPCR. C: CCL22 protein analyzed by ELISA assay. ****, P<0.001.

Article Snippet: The concentration of CCL22 in the supernatant was tested according to the instructions of ELISA kit operation (CUSABIO, CSB-E04660h).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: LncRNA HOTAIR promotes proliferation, invasion and migration in NSCLC cells via the CCL22 signaling pathway

doi: 10.1371/journal.pone.0263997

Figure Lengend Snippet: A: The HOTAIR and CCL22 mRNA expression examined by RT-qPCR B: Cell apoptosis detected by Flow cytometry. C: The cell proliferation tested by MTS assay. D: The CCL22 protein expression tested by ELISA. E: The cell migration and invasion detected by Transwell assay. **, P<0.05; ***, P<0.01; ****, P<0.001;ns, no significance.

Article Snippet: The concentration of CCL22 in the supernatant was tested according to the instructions of ELISA kit operation (CUSABIO, CSB-E04660h).

Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry, MTS Assay, Enzyme-linked Immunosorbent Assay, Migration, Transwell Assay

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: TNFAIP8 expression is upregulated in BCC, SCC, and melanoma patient skin tissues. ( A ) Relative immunohistochemical TNFAIP8 expression in indicated skin cancer tissues is presented. ( B ) Immunohistochemical expression levels of TNFAIP8 protein from normal skin (n = 6), BCC (n = 6), H-SCC (n = 4), M-SCC (n = 4), L-SCC (n = 4), nevus (n = 6), and melanoma (n = 6) tissues were quantified using ImageJ software ( https://imagej.nih.gov/ij/ ) and plotted. The data represent mean ± SEM from 4 to 6 skin cancer tissues. * P < 0.05 relative to normal skin tissues. ( C ) Expression of TNFAIP8 transcripts between cutaneous melanoma (n = 45) and melanoma precursor (n = 18) samples were analyzed and presented from the Oncomine dataset ( https://www.oncomine.org/resource/login.html accessed October 17, 2020). *** P < 0.001 relatives to cutaneous melanoma tissues. ( D , E ) Expression of wild-type B-RAF, mutant B-RAF V600E , and TNFAIP8 in HaCaT, SK-MEL-2, A431, A375, and A2058 cells were analyzed by western blotting. Fifty micrograms of lysates from normal and skin cancer cells were immunoblotted with B-RAF, TNFAIP8, GAPDH, and β-actin antibodies. Immunoreactive bands were visualized using ECL chemiluminescence detection reagents after exposing the blots on X-ray films ( D ) or by scanning the blots with an Odyssey CLx Imager ( E ). The immunoblot scans were converted into grayscale and presented. ( F ) Expression of TNFAIP8 mRNA from indicated normal skin cells and skin cancer cells were analyzed by RT/qPCR as described in the “ ” section.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: Expressing, Immunohistochemical staining, Software, Mutagenesis, Western Blot, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: TNFα-induced TNFAIP8 expression in skin cancer cells ( A ) Schematic represents TNFAIP8 isoform-specific forward and reverse primer design. ( B ) The expression of different variants/isoforms of TNFAIP8 in normal HaCaT and skin cancer cells was analyzed by RT-PCR. NC–negative control (no cDNA). ( C ) HaCaT, A431, A375, and A2058 cells were treated with vehicle or TNFα (10–50 ng/ml) for 30 h, and cell lysates were immunoblotted with TNFAIP8 or β-actin antibodies. Immunoreactive bands were visualized using ECL chemiluminescence detection reagents and the blots were scanned using an Odyssey CLx imager. The immunoblot scans were converted into grayscale and presented. ( D ) Similarly, normal and skin cancer cells were treated with vehicle or TNFα (10-50 ng/ml) for 30 h, and TNFAIP8 mRNA expression was analyzed by RT/qPCR as described in the “ ” section. * P < 0.05, ** P < 0.01, *** P < 0.001 relative to control/vehicle-treated cells. ( E ) HaCaT, A431, A375 and, A2058 cells were treated with vehicle or TNFα (10–50 ng/ml) for 48 h and, relative cell survival was analyzed by MTT assay as described in the “ ” section. Results are representative of three independent experiments. * P < 0.05, *** P < 0.001 relative to control/vehicle-treated cells.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Western Blot, Quantitative RT-PCR, Control, MTT Assay

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: TNFAIP8 regulates cell proliferation and migration in skin cancer cells. ( A ) Western blotting analysis of TNFAIP8-Myc tagged protein expression in HaCaT and skin cancer cells. Immunoreactive bands were developed using ECL chemiluminescence detection reagents and the blots were scanned using an Odyssey CLx imager. The immunoblot scans were converted into grayscale and presented. (B) Effect of TNFAIP8-Myc protein overexpression on HaCaT, A431, A375 and, A2058 on cell survival was measured by MTT assay. Results are representative of three independent experiments. * P < 0.05 relative to EV transfected cells. ( C ) The effect of overexpression of TNFAIP8-Myc on cell colony formation was analyzed and plotted (upper and lower panels) as described in the “ ” section. * P < 0.05 relative to EV transfected cells. ( D ) HaCaT, A431, A375 and, A2058 cells were transfected with control siRNA or TNFAIP8 siRNA (100 nM) for 30 h, and cell lysates were western blotted with anti-TNFAIP8 and β-actin antibodies. Western blots were developed using ECL chemiluminescence detection reagents and the blots were scanned using an Odyssey CLx imager. The immunoblot scans were converted into grayscale and presented. TNFAIP8 and β-actin protein levels were quantified using ImageJ software ( https://imagej.nih.gov/ij/ ) and presented below. ( E ) The effect of TNFAIP8 knockdown by siRNA on cell survival was analyzed by MTT assay. The experiments were independently performed three times. * P < 0.05 relative to control siRNA transfected cells. ( F ) Control siRNA and TNFAIP8 siRNA-transfected HaCaT, A431, A375 and A2058 cells (3000 cells/well) were re-plated in 6-well plates in triplicate for 7–10 days and skin cancer cell colony formation was analyzed and plotted (upper and lower panels) as described in the “ ” section. * P < 0.05 relative to control siRNA transfected cells. ( G ) Wound healing assay: HaCaT, A431, A375 and A2058 cells were transfected with control siRNA or TNFAIP8 siRNA for 48 h. The effect of the TNFAIP8 knockdown on cell migration was analyzed using the wound healing assay. Representative images of the wound healing assay (top) and the calculated scratch area (bottom) are shown. * P < 0.05 relative to control siRNA transfected cells.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: Migration, Western Blot, Expressing, Over Expression, MTT Assay, Transfection, Control, Software, Knockdown, Wound Healing Assay

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: miR-205-5p targets the 3′UTR of TNFAIP8 and inhibits TNFAIP8 expression. ( A ) The binding site of miR-205-5p in the 3′UTR of TNFAIP8 was analyzed by TargetScan ( http://www.targetscan.org/vert_72/ ) and presented. We also generated a wild type and mutated binding nucleotides of miR-205-5p to the 3′UTR of TNFAIP8. Wild-type miR-205-5p and mutant miR-205-5p mimics are shown. ( B ) Relative expression of miR-205-5p in HaCaT, A431, A375, A2058, and SK-MEL-2 cell lines was analyzed by RT/qPCR as described in the “ ” section. ( C ) Luciferase reporter assay: A431 and A2058 cells were transfected with TNFAIP8-3′UTR-Luciferase reporter construct (0.5 µg) for 18 h and then transfected with NC mimic or wild-type miR-205-5p mimic or mutant miR-205-5p mimic for an additional 24 h. Transfected cells were lysed, and luciferase activity was measured. Results are representative of two independent experiments. ** P < 0.01, *** P < 0.001 compared with NC or mutant-type miR-205-5p mimic. ( D , E ) Normal or skin cancer cells were transfected with NC or miR-205-5p mimic for 48 h and the overexpression of miR-205-5p mimic and TNFAIP8 transcripts were analyzed by RT/qPCR. *** P < 0.001 compared with NC-transfected cells. * P < 0.05 compared with NC-transfected cells. ns-no significance. ( F ) HaCaT, A431 and A2058 cell lines were transfected with NC mimic or miR-205-5p mimic for 40 h and expression of endogenous TNFAIP8 protein were analyzed by western blotting. Western blots were developed using ECL chemiluminescence detection reagents, the blots were exposed to X-Ray films, scans were converted into grayscale and presented. ( G ) HaCaT, 431 and A2058 cell lines were grown on a coverslip and transfected with Cy3-labeled NC mimic or Cy3-labeled miR-205-5p mimic and the effect of Cy3-labeled miR-205-5p mimic on endogenous TNFAIP8 expression (green) was visualized by immunofluorescence as described in the “ ” section.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: Expressing, Binding Assay, Generated, Mutagenesis, Quantitative RT-PCR, Luciferase, Reporter Assay, Transfection, Construct, Activity Assay, Over Expression, Western Blot, Labeling, Immunofluorescence

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: miR-205-5p targets TNFAIP8 and reduces TNFAIP8-mediated cell proliferation and autophagy. ( A ) HaCaT, A431 and A2058 cells were transfected with NC mimic or miR-205-5p mimic for 48 h and relative cell survival was analyzed by MTT assay. The experiments were independently repeated three times. * P < 0.05 compared with NC mimic transfected cells. ( B ) NC mimic or miR-205-5p mimic transfected cells were re-plated in 6-well plates for 7–10 days and cell colony formation was analyzed and plotted (upper and lower panels) as described in the “ ” section. The experiment was repeated twice in triplicates. * P < 0.05 relative to NC mimic transfected cells. ( C ) HaCaT, A431 and A2058 cells were transfected EV or TNFAIP8-Myc plasmids for 30 h and lysates were western blotted with anti-Myc, anti-LC3β I/II, and anti-β-actin antibodies. Western blots were developed using ECL chemiluminescence detection reagents, the blots were exposed on X-ray films, converted into grayscale, and presented. ( D ) HaCaT, lanoma A431 and A2058 cells were transfected NC mimic or miR-205-5p mimic for 30 h and lysates were western blotted with anti-TNFAIP8, anti-LC3β I/II, anti-p62, and anti-β-actin antibodies. Western blots were developed using ECL chemiluminescence detection reagents, exposed on X-Ray films, converted into grayscale, and presented. ( E ) HaCaT, A431 and A2058 cells were grown on a coverslip transfected with Cy3-labeled NC mimic or Cy3-labeled miR-205-5p mimic and the effect of Cy3-labeled miR-205-5p mimic on endogenous LC3β I/II expression (green) and related puncta formation were visualized by immunofluorescence (upper panels) as described in the “ ” section (upper panel). LC3β I/II and related puncta were measured from individual cells (n = 10) and plotted (lower panels). ** P < 0.01, *** P < 0.001 relatives to Cy3 labeled-NC mimic transfected cells.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: Transfection, MTT Assay, Western Blot, Labeling, Expressing, Immunofluorescence

Journal: Scientific Reports

Article Title: MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8

doi: 10.1038/s41598-021-85097-6

Figure Lengend Snippet: miR-205-5p targets TNFAIP8 in skin cancer cells and increase sensitivity to vemurafenib. ( A ) HaCaT cells, and skin cancer SK-MEL-2, A375 and A2058 cell lines were treated with increasing concentrations of vemurafenib for 48 h and relative cell survival/proliferation was measured by MTT assay. The experiments were independently repeated three times. ** P < 0.01, *** P < 0.001 compared to vehicle-treated cells. ( B ) HaCaT cells and skin cancer SK-MEL-2, A375 and A2058 cell lines were transfected with NC mimic or miR-205-5p mimic for 24 h and treated with vemurafenib or vehicle for an additional 48 h and relative cell survival were measured by MTT assay as described in the “ ” section. * P < 0.05, ** P < 0.01, *** P < 0.001 compared to NC mimic vehicle-treated cells or NC mimic and vemurafenib-treated cells. ns-no significance. ( C ) Melanoma A375 cells were transfected with NC mimic or miR-205-5p mimic for 18 h and then cells were treated with vemurafenib for an additional 24 h. Fifty micrograms of lysates were immunoblotted with anti-cleaved-PARP and anti-β-tubulin antibodies. Western blots were developed using ECL chemiluminescence detection reagents, the blots were scanned using Azure C-500 Biosystem, scans were converted into grayscale and presented. ( D ) Melanoma A375 cells were transfected with EV of TNFAIP8-Myc plasmid for 18 h first and then cells were treated with vemurafenib or paclitaxel for 24 h as indicated and fifty micrograms lysates were immunoblotted with anti-Myc, anti-cleaved-PARP, and anti-β-actin antibodies. Western blots were developed using ECL chemiluminescence detection reagents and the blots were scanned using an Odyssey CLx imager. The immunoblot scans were converted into grayscale and presented. ( E ) Fifty micrograms of lysates from A375 and A2058 cells or A375R and A2058R (vemurafenib resistant) cells were immunoblotted with anti-TNFAIP8, anti-LC3β I/II, and anti-β-actin antibodies. Western blots were developed using ECL chemiluminescence detection reagents, the blots were exposed on X-Ray films, scans were converted into grayscale and presented. ( F ) Relative cell proliferation/cell survival rate of A375 and A2058 cells, or A375R and A2058R (vemurafenib resistant) cells were measured by using WST-1 reagent (Cayman Chemical) (upper panel) or MTT assay reagent (lower panel) and plotted. ** P < 0.01, *** P < 0.001 compared to A375 and A2058 parent cells. Results are representative of three independent experiments. Vem. Vemurafenib.

Article Snippet: Anti-β-actin antibody from Sigma (St. Louis, MO) and anti-TNFAIP8 antibody from Proteintech Group (Rosemont, IL) was also purchased.

Techniques: MTT Assay, Transfection, Western Blot, Plasmid Preparation