Journal: Oncotarget

Article Title: Role of SATB2 in human pancreatic cancer: Implications in transformation and a promising biomarker

doi: 10.18632/oncotarget.10860

Figure Lengend Snippet: ( A ) Protein expression of SATB2 in human pancreatic normal ductal epithelial cells (HPNE), pancreatic cancer cell lines and Pan CSCs. Crude proteins were isolated and the expression of SATB2 was measured by the Western blot analysis. β-actin was used as a loading control. ( B ) Expression of SATB2 mRNA in HPNE, pancreatic cancer cell lines and Pan CSCs. RNA was isolated and the expression of SATB2 was measured by qRT-PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, #, & and % = significantly different from HPNE ( P < 0.05). ND = Not Detected. Pan CSCs = Pancreatic Cancer Stem Cells.

Article Snippet: Antibodies against SATB2 and β-actin were purchased from Abcam (Cambridge, MA).

Techniques: Expressing, Isolation, Western Blot, Control, Quantitative RT-PCR

Journal: Oncotarget

Article Title: Role of SATB2 in human pancreatic cancer: Implications in transformation and a promising biomarker

doi: 10.18632/oncotarget.10860

Figure Lengend Snippet: ( A ) HPNE cells were stably transduced with lentiviral particles expressing either empty vector or SATB2 cDNA. SATB2 expression was measured by the Western blot analysis, RT-PCR and immunocytochemistry. ( B ), Proliferation of HPNE/Empty Vector and HPNE/SATB2 cDNA cells was measured for 6 days. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from respective empty vector groups, P < 0.05. ( C ), Colony and spheroid formation. Colony formation in soft agar and spheroid formation in suspension of HPNE/Empty Vector and HPNE/SATB2 cDNA cells were measured. ( D ) Upper panel, RNA expression of stem cell markers. RNA was isolated and the expression of stem cell markers (CD133 and CD44) was measured by qRT-PCR analysis. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( P < 0.05). Gene expression of HPNE/Empty Vector cells was normalized to 1. Lower panel, Protein expression of stem cell markers. Cell lysates were collected from HPNE/Empty Vector and HPNE/SATB2 cDNA cells, and the expression of CD24, CD44 and CD133 was measured by the Western blot analysis. β-actin was used as a loading control. ( E ) Upper panel, RNA was isolated and the expression of transcription factors (c-Myc, Nanog and Oct-4) was measured by qRT-PCR analysis. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( P < 0.05). Gene expression of HPNE/Empty Vector cells was normalized to 1. Lower panel, Protein expression of c-Myc, Nanog and Oct-4 Cell lysates were collected from HPNE/Empty Vector and HPNE/SATB2 cDNA cells, and the expression of c-Myc, Nanog and Oct-4 was measured by the Western blot analysis. β-actin was used as a loading control. ( F ), Upper panel, RNA was isolated and the expression of Bcl-2 and XIAP was measured by qRT-PCR analysis. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( P < 0.05). Gene expression of HPNE/Empty Vector cells was normalized to 1. Lower panel, Protein expression of Bcl-2. Cell lysates were collected from HPNE/Empty Vector and HPNE/SATB2 cDNA cells, and the expression of Bcl-2 was measured by the Western blot analysis. β-actin was used as a loading control.

Article Snippet: Antibodies against SATB2 and β-actin were purchased from Abcam (Cambridge, MA).

Techniques: Stable Transfection, Transduction, Expressing, Plasmid Preparation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry, Suspension, RNA Expression, Isolation, Quantitative RT-PCR, Gene Expression, Control

Journal: Oncotarget

Article Title: Role of SATB2 in human pancreatic cancer: Implications in transformation and a promising biomarker

doi: 10.18632/oncotarget.10860

Figure Lengend Snippet: ( A ) Cell Motility assay. HPNE/Empty Vector and HPNE/SATB2 cDNA cells were grown in petri dishes. After cell attachment, scratch lines were made with the fine pipette tips in both the groups. Phase contrast images of cells were captured at 0 h and 24 h time points. ( B ) Transwell Migration Assay. 1 × 10 5 cells were plated on to the top chamber of the noncoated membrane (24-well insert; pore size, 8 μm; Corning Costar) and allowed to migrate toward serum-containing medium in the lower chamber. Cells were fixed after 24 hours of incubation with methanol and stained with Diff-Quick Fixative Solutions. Data represent mean ( n = 4) ± SD. * = significantly different at P < 0.05. ( C ) Transwell Invasion Assay. 1 × 105 cells were plated on to the top chamber of the Matrigel coated Membrane. After 48 hours, Matrigel-coated inserts were fixed and stained. The number of cells invading through the membrane was counted. Data represent mean ± SD. * = significantly different at P < 0.05. ( D ) Expression of E-Cadherin, N-cadherin and Zeb1 in HPNE/Empty Vector and HPNE/SATB2 cDNA cells. RNA was isolated and the expression of E-Cadherin, N-cadherin and Zeb1 was measured by qRT-PCR analysis. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE/Empty Vector group ( P < 0.05). Gene expression of HPNE/Empty Vector group was normalized to 1. ( E ), Protein expression of E-Cadherin, N-cadherin and Zeb1. Cell lysates were collected from HPNE/Empty Vector and HPNE/SATB2 cDNA cells, and the expression of E-Cadherin, N-cadherin and Zeb1 was measured by the Western blot analysis. β-actin was used as a loading control.

Article Snippet: Antibodies against SATB2 and β-actin were purchased from Abcam (Cambridge, MA).

Techniques: Motility Assay, Plasmid Preparation, Cell Attachment Assay, Transferring, Transwell Migration Assay, Membrane, Pore Size, Incubation, Staining, Diff-Quik, Transwell Invasion Assay, Expressing, Isolation, Quantitative RT-PCR, Gene Expression, Western Blot, Control

Journal: Oncotarget

Article Title: Role of SATB2 in human pancreatic cancer: Implications in transformation and a promising biomarker

doi: 10.18632/oncotarget.10860

Figure Lengend Snippet: ( A ) Expression of SATB2. PANC-1, AsPC-1 and Pan CSCs were transduced with either scrambled or SATB2 shRNA. Cell lysates were collected and the expression of SATB2 was measured by the Western blot analysis. β-actin was used as a loading control. ( B ) Cell proliferation of PANC-1/Scrambled, PANC-1/SATB2 shRNA, AsPC-1/Scrambled, AsPC-1/SATB2 shRNA, Pan CSCs/Scrambled and Pan CSCs/SATB2 shRNA groups was measured over 6-day period. ( C ) Colony formation Assay. PANC-1/Scrambled, PANC-1/SATB2 shRNA, AsPC-1/Scrambled, AsPC-1/SATB2 shRNA, Pan CSCs/Scrambled and Pan CSCs/SATB2 shRNA cells were seeded, and number of colonies formed at 21 days were counted. Data represent mean ( n = 4) ± SD. * = significantly different from HPNE empty vector ( P < 0.05).

Article Snippet: Antibodies against SATB2 and β-actin were purchased from Abcam (Cambridge, MA).

Techniques: Expressing, Transduction, shRNA, Western Blot, Control, Colony Assay, Plasmid Preparation

Journal: Oncotarget

Article Title: Role of SATB2 in human pancreatic cancer: Implications in transformation and a promising biomarker

doi: 10.18632/oncotarget.10860

Figure Lengend Snippet: ( A ) Cell Motility Assay. Pan CSCs/Scrambled, Pan CSCs/SATB2 shRNA, AsPC-1/Scrambled, AsPC-1/SATB2 shRNA, PANC-1/Scrambled, PANC-1/SATB2 shRNA cells were grown in petri dishes. After 18 hours of incubation, cells were scratched with the fine pipette tips. Phase contrast images of scratched cells were captured at 0 h, 24 h and 48 h time points. ( B ) Transwell Migration Assay. Transwell migration assay was performed in Pan CSCs/Scrambled, and Pan CSCs/SATB2 shRNA cells as described in Materials and Methods. Data represent mean ± SD. * = significantly different at P < 0.05. ( C ) Transwell Invasion Assay. Transwell invasion assay was performed in Pan CSCs/Scrambled and Pan CSCs/SATB2 shRNA as described in Materials and Methods. Data represent mean ± SD. * = significantly different at P < 0.05. ( D ) Expression of EMT-related genes/proteins. Upper panel, RNA was isolated and the expression of E-cadherin, N-cadherin, and Zeb1 in Pan CSCs/Scrambled, and Pan CSCs/SATB2 shRNA cells was measured by qRT-PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different between groups ( P < 0.05). Lower panel, Protein expression of E-Cadherin, N-cadherin, Zeb1 and Snail. Cell lysates were collected, and the expression of E-Cadherin, N-cadherin, Zeb1 and Snail was measured by the Western blot analysis. β-Actin was used as a loading control. ( E ) Expression of cell proliferation/survival genes/proteins. Upper panel, RNA was isolated and the expression of Bcl-2 and XIAP in Pan CSCs/Scrambled, and Pan CSCs/SATB2 shRNA cells was measured by qRT-PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different between groups ( P < 0.05). Lower panel, Protein expression of Bcl-2 and XIAP. Cell lysates were collected, and the expression of Bcl-2 and XIAP was measured by the Western blot analysis. β-Actin was used as a loading control. ( F ) Expression of pluripotency maintaining factors. Upper panel, RNA was isolated and the expression of c-Myc, Nanog, and Oct-4 in Pan CSCs/Scrambled, and Pan CSCs/SATB2 shRNA cells was measured by qRT-PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different between groups ( P < 0.05). Lower panel, Protein expression of c-Myc, Nanog, and Oct-4. Cell lysates were collected, and the expression of c-Myc, Nanog, and Oct-4 was measured by the Western blot analysis. β-Actin was used as a loading control. ( G ) Expression of stem cell markers. Upper panel, RNA was isolated and the expression of CD44 in Pan CSCs/Scrambled, and Pan CSCs/SATB2 shRNA cells was measured by qRT-PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. * = significantly different between groups ( P < 0.05). Lower panel, Protein expression of stem cell markers. Cell lysates were collected, and the expression of CD24, CD44 and CD133 was measured by the Western blot analysis. β-Actin was used as a loading control.

Article Snippet: Antibodies against SATB2 and β-actin were purchased from Abcam (Cambridge, MA).

Techniques: Motility Assay, shRNA, Incubation, Transferring, Transwell Migration Assay, Transwell Invasion Assay, Expressing, Isolation, Quantitative RT-PCR, Control, Western Blot

Journal: Endocrinology

Article Title: Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.

doi: 10.1210/en.2011-1926

Figure Lengend Snippet: FIG. 2. Ex vivo analysis of hPRL gene promoter activation. A, qPCR of hPRL promoter activity of PE , BM cells, and PBMC in T24 and S24 rats. A minimum of four samples from each treatment group was assayed in duplicate. Data are presented as mean SEM; *, P 0.05. B, Luciferase activity of PE and BM cells isolated from S24 and T24 rats. ***, P 0.001. C, Ex vivo bioluminescent imaging of FITC-labeled, CD11b adherent cells from the PE, BM, and PBMC of S24 transgenic rats. Scale bar, 50 m. D, Comparison of bioluminescence between PE cells from a S24 rat and a T24 rat. S24, saline 24 h; T24, TG 24 h; RLU, Relative luminescence units. Scale bar, 100 m.

Article Snippet: Adherent cells were blocked with a mouse antirat CD32 antibody (BD Pharmingen, San Diego, CA), then stained with an antirat CD11b fluorescein isothiocyanate (FITC)-conjugated antibody or matched isotype control (1:10 antibody; AbD Serotec, Kidlington, UK).

Techniques: Ex Vivo, Activation Assay, Activity Assay, Luciferase, Isolation, Imaging, Labeling, Transgenic Assay, Comparison, Saline

Journal: Endocrinology

Article Title: Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.

doi: 10.1210/en.2011-1926

Figure Lengend Snippet: FIG. 3. Identification of subpopulations of CD11b cells expressing luciferase in the PE. A, FACS analysis of cells isolated from PE of S24 rats: two-dimensional dot plot (ssc vs. fsc) showing two populations of CD11b-Alexa Fluor 647-stained cells (histograms, central panel). Blue, CD11b/sschigh; green, CD11B/ssclow. Diff-Quick staining of the corresponding sorted cell populations (right panels). Small black arrow, Mast cell, dark purple granules; small white arrow, eosinophils, red granules; large white arrow, B-cell, small cell with little cytoplasm; large black arrow, macrophage. Scale bar, 50 m. B, qPCR quantification of luciferase transcript in the S24 CD11b/ssclow population (n 5). C, FACS analysis of cells isolated from PE of T24 rats. Two-dimensional dot plot (ssc vs. fsc), representing three populations of CD11b-Alexa Fluor 647- and/or Gra-PE-stained cells (histograms, central panel). Blue, CD11b/sschigh; green, CD11b/Gra; pink, CD11b/Gra. Diff-Quick staining of the corresponding sorted cell populations (right panels). Scale bar, 50 m. D, qPCR quantification of luciferase transcript in the T24 CD11b/Gra (n 3) and CD11b/Gra (n 5) populations. E, RT-PCR for detection of extrapituitary luciferase and endogenous rPRL gene expression. In addition to the expected 122-bp product, a larger band (260 bp) was produced by alternative splicing as described previously (31). M, marker; 1, Luciferase from T24 CD11b/Gra; 2, rPRL from T24 CD11b/Gra; 3, Luciferase from T24 CD11b/Gra; 4, rPRL from T24 CD11b/Gra; 5, rPRL on pituitary (control); S24, saline 24 h; T24, TG 24 h.

Article Snippet: Adherent cells were blocked with a mouse antirat CD32 antibody (BD Pharmingen, San Diego, CA), then stained with an antirat CD11b fluorescein isothiocyanate (FITC)-conjugated antibody or matched isotype control (1:10 antibody; AbD Serotec, Kidlington, UK).

Techniques: Expressing, Luciferase, Isolation, Staining, Diff-Quik, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Produced, Alternative Splicing, Marker, Control, Saline

Journal: Endocrinology

Article Title: Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.

doi: 10.1210/en.2011-1926

Figure Lengend Snippet: FIG. 4. Subpopulations of CD11b cells expressing luciferase in BM and PBMC. A, FACS analysis of cells isolated from BM of T24 rats. Two- dimensional dot plot (ssc vs. fsc) showing two populations of CD11b-Alexa Fluor 647- and/or Gra-PE-stained cells (histograms, central panel). Blue, CD11b/sschigh (Gra); green, CD11b/ssclow (Gra). Diff-Quick staining of corresponding sorted cell populations (right panels). Black arrow, Monocytes; white arrow, neutrophils. Scale bar, 50 m. B, qPCR quantification of luciferase transcript in the S24 (n 5) and T24 (n 6) CD11b/ssclow population in BM. C, RT-PCR for the detection of extrapituitary luciferase and endogenous rPRL gene expression. M, Marker; 1 and 3, Luciferase on T24 CD11b/sschigh; 2 and 4, rPRL on T24 CD11b/sschigh; 5, rPRL on pituitary (control). D, FACS analysis of cells isolated from PB of T24 rats. Two-dimensional dot plot (ssc vs. fsc) showing two populations of CD11b-Alexa Fluor 647- and/or Gra-PE-stained cells (histograms, central panel). Blue, CD11b/Gra and green, CD11b/Gra. E, qPCR quantification of luciferase transcript in the S24 (n 5) and T24 (n 5) CD11b/Gra and T24 CD11b/Gra (n 5 in each group). S24, saline 24 h; T24, TG 24 h.

Article Snippet: Adherent cells were blocked with a mouse antirat CD32 antibody (BD Pharmingen, San Diego, CA), then stained with an antirat CD11b fluorescein isothiocyanate (FITC)-conjugated antibody or matched isotype control (1:10 antibody; AbD Serotec, Kidlington, UK).

Techniques: Expressing, Luciferase, Isolation, Staining, Diff-Quik, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Marker, Control, Saline

Journal: Endocrinology

Article Title: Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.

doi: 10.1210/en.2011-1926

Figure Lengend Snippet: FIG. 7. Ex vivo stimulation of PRL expression in human peripheral blood monocytes. A, Peripheral blood monocytes stained with CD11b antibody (left panel). Diff-Quick stain of CD11b cells to confirm monocytes morphology. Scale bar, 50 m. B and C, RT-PCR analysis of two distinct donors, showing the bad pattern of extrapituitary hPRL gene expression. D and E, qPCR of hPRL extrapituitary gene expression in human CD11b PBMC of two donors. Cells were treated with TNF- (TNF), LPS, and TG for 16 h, then analyzed for hPRL gene expression. CycloA, Cyclophilin A; Ctrl, no-stimulation control.

Article Snippet: Adherent cells were blocked with a mouse antirat CD32 antibody (BD Pharmingen, San Diego, CA), then stained with an antirat CD11b fluorescein isothiocyanate (FITC)-conjugated antibody or matched isotype control (1:10 antibody; AbD Serotec, Kidlington, UK).

Techniques: Ex Vivo, Expressing, Staining, Diff-Quik, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Control

Journal: Theranostics

Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

doi: 10.7150/thno.72269

Figure Lengend Snippet: SH2 superbinder exhibited excellent security by targeting lung fibroblasts based on pY levels in vivo. A , Schematic showing progression of SH2 superbinder treatment in control mice. B , Percent survival during last 7 days after SH2 superbinder injection (n = 8). C , HE staining after SH2 superbinder challenge. Images showing the panoramic and partial view of lungs. D , Neutrophil accumulation measured by MPO assay in lung tissues after GST, GST-SH2 WT, GST-SH2 TrM or SH2-TrM treatment in saline groups (n = 6). E , Total cell counting in BALF (n = 6). F-G , Changes in different kinds of cells by Giemsa and Diff-quick staining in BALF (n = 6). H , Inflammatory cytokines in BALF measured by ELISA (n = 6). I , The determination of liver function index in serum (n = 6). J , Schematic showing progression of SH2 superbinder treatment in BLM-treated mice. K , Western blot of GST tag of different tissues in BLM group treated with GST-SH2 TrM. L , The process of FACS for mice lungs. M , Western blot of pY levels in different lung cells which were isolated by FACS of BLM treated 14 days mice. N , The fluorescence images of SH2 superbinder entering into activated fibroblasts, epithelial cells (EpCAM + ), leukocytes (CD45 + ) and endothelial cells (CD31 + ). O-R , Representative images of SH2 superbinder in myofibroblast (α-SMA + ) ( O ), epithelial cells (EpCAM + ) ( P ), leukocytes (CD45 + ) ( Q ) and endothelial cells (CD31 + ) ( R ) of BLM-treated mice.

Article Snippet: The amounts of IL-1β, IL-6, IL-10 and TNF-α in BALF of mice were measured by using mouse ELISA kits from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vivo, Control, Injection, Staining, MPO Assay, Saline, Cell Counting, Diff-Quik, Enzyme-linked Immunosorbent Assay, Western Blot, Isolation, Fluorescence

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: TNFα is correlated with human oral cancer pain scores. ( a ) TNFα protein concentration is higher in cancer tissues compared to anatomically matched contralateral healthy tissues from the same patient (n = 10, * P < 0.05, paired t-test). ( b ) Patients were asked to answer the Oral Cancer Pain Questionnaire before surgery. The mean pain score from patients correlated positively with percentage change in TNFα concentration between cancer and matched contralateral normal tissues ( r = 0.7, P < 0.05).

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Protein Concentration, Concentration Assay

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Blocking TNFα or JNK inhibits nociception in mice with cancer. ( a ) After 16 weeks of 4NQO treatment, mice exhibited significant increase in gnaw-time from its respective baseline (pre-injection). Propylene glycol (PG) treatment did not affect gnaw-time. In 4NQO tongue cancer mice, C-87 (12.5 mg/kg) IP injection significantly reduced percentage of gnaw-time change from baseline (n = 8) 1 h post-injection than the vehicle (10% DMSO) treated cancer mice (n = 5). C-87 (n = 6) or vehicle (n = 4) had no effect in non-cancer mice treated with PG alone. ( b ) Mice with paw SCC developed cancer pain at PID7. C-87 and the JNK inhibitor SP600125 treatment significantly reduced mechanical nociception compared to vehicle at 1, 3, and 6 h after treatment compared to the control group. 24 h after the treatment the analgesic effect of C-87 was gone (n = 5 per group, Two-way ANOVA). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Blocking Assay, Injection, Control

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Blocking TNFα inhibits cancer cell growth, migration, and cytokine release. ( a ) Growth rate, measured with the RTCA, following different doses of C-87 treatment in HSC-3 cell culture. C-87 inhibited oral cancer cell growth in a dose dependent manner. One-way ANOVA with Tukey's post hoc analysis. ( b ) Mice with C-87 treatment (n = 7) exhibited a significant decrease in the paw volume compared to the vehicle control mice (n = 6) at PID14, 18, and 21 (two-way ANOVA). Arrow indicates C-87 injection. ( c ) C-87 treated paw cancer mice (n = 6) had smaller tumor area relative to the total paw area compared to vehicle treated paw cancer mice (n = 4). Tumor areas and total paw areas were quantified using H&E stained paw sections. Mann–Whitney U-test. ( d ) Representative H&E stained pictures showing a normal mouse paw, a cancer mouse paw, and a cancer paw treated with C-87 (10 × inset). Scale bar: 100 μm. Images were taken and quantified using Nikon imaging software NIS-Elements F Ver4.60.00. ( e ) C-87 treatment reduced the concentration of TNFα, NGF, IL1β, IL4, MIP3α, IL28β, and IL33 in the paw tumor. Data were presented as fold change of cytokines/chemokines measured from tumor paws over normal paws. n = 6 per group. Mann–Whitney U test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Blocking Assay, Migration, Cell Culture, Control, Injection, Staining, MANN-WHITNEY, Imaging, Software, Concentration Assay

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: TNFα mediates Schwann cell proliferation and migration in vitro. ( a ) The presence of either DOK or HSC-3 cells in cell inserts increased Schwann cell proliferation 48 h following co-culture in the MTS assay. Increased Schwann cell proliferation induced by the presence of HSC-3 cells was inhibited by adding C-87 into inserts. Representative images of cells with Hoechst stain were shown under each culture condition. OD: optical density. ( b ) Schwann cells are more migratory in the presence of HSC-3 cells compared to the DMEM control while DOK reduced Schwann cell migration. Adding C-87 into the HSC-3 culture reduced Schwann cell migration. ( c ) Adding TNFα to the media at the bottom chamber increased Schwann cells migration compared to the DMEM control. Neutralizing TNFα with C-87 decreased Schwann cell migration. ( d ) Schwann cells induced increased HSC-3 cell migration compared to the DMEM control; adding C-87 (20 µM) into the Schwann cell culture in the bottom chamber blocked this increase. ( b – d ), images shown are representative diff-quick stained migrated cells. a-d, one-way ANOVA with Tukey's post hoc analysis. SCs: Schwann cells. Scale bar: 100 μm. * P < 0.05; ** P < 0.01; *** P < 0.001. Images were taken using Nikon imaging software NIS-Elements F Ver4.60.00.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Migration, In Vitro, Co-Culture Assay, MTS Assay, Staining, Control, Cell Culture, Diff-Quik, Imaging, Software

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: The effect TNFα on the expression of Schwann cell activation markers in vitro. TNFα treatment increased c-Jun ( a , b ), GFAP ( c , d ), and p75 ( e – f ) immunofluorescence intensity and protein expression in cultured Schwann cells compared to the DMEM control. TNFα treatment decreased MBP immunofluorescence intensity and protein expression in cultured Schwann cells compared to the DMEM control ( g – h ). Full-length gel blots were provided in the Supplemental Fig. online. Scale bar: 100 μm. Student’s t-test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Expressing, Activation Assay, In Vitro, Immunofluorescence, Cell Culture, Control

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Activated Schwann cells release increased NGF and TNFα. ( a ) Schwann cells co-cultured with HSC-3 cells overexpressed c-Jun, GFAP, p75 but downregulated MBP compared to control Schwann cells (media alone). Full-length gel blots were provided in Supplemental Fig. online. ( b ) Both DOK and HSC-3 co-cultures increased TNFα mRNA expression in Schwann cells compared to control Schwann cells. ( c ) TNFα protein concentration in Schwann cells co-cultured with either DOK or HSC-3 cells compared to control Schwann cells. ( d ) HSC-3 cell or DRK co-culture increased NGF release in Schwann cells compared with control Schwann cells. ( e ) Adding TNFα in cell culture media stimulated increased NGF release compared with control Schwann cells. One-way ANOVA. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Cell Culture, Control, Expressing, Protein Concentration, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Anti-Warburg Mechanism of Ginsenoside F2 in Human Cervical Cancer Cells via Activation of miR193a-5p and Inhibition of β-Catenin/c-Myc/Hexokinase 2 Signaling Axis

doi: 10.3390/ijms25179418

Figure Lengend Snippet: Effect of GF2 on cytotoxicity and colony formation in HeLa and SiHa cells. ( A ) Chemical structure of GF2 (MW = 785.03). ( B ) Cytotoxic effect of GF2 in HeLa and SiHa cells. The cells were exposed to various concentrations of GF2 for 24 h, and cell viability was assessed by MTT assay. * p < 0.05, ** p < 0.01, and *** p < 0.001 versus the untreated control. ( C ) Antiproliferative effect of GF2 in HeLa and SiHa cells. HeLa and SiHa cells were treated with GF2 (0, 50, and 70 μM), and then colony formation took place over 1 week. The colonies were visualized by staining with Diff-Quick solution (Sysmex, Kobe, Japan). The data represent the means ± SD.

Article Snippet: HeLa (ccl-2, ATCC, Manassas, VA, USA) and SiHa (HTB-35, ATCC, Manassas, VA, USA) cervical cancer cells supplied by the ATCC company were cultured in Dulbecco’s modified Eagle medium (DMEM), including 10% FBS and 1% antibiotic antimycotic solution (Welgene, Republic of Korea).

Techniques: MTT Assay, Control, Staining, Diff-Quik

Journal: International Journal of Molecular Sciences

Article Title: Anti-Warburg Mechanism of Ginsenoside F2 in Human Cervical Cancer Cells via Activation of miR193a-5p and Inhibition of β-Catenin/c-Myc/Hexokinase 2 Signaling Axis

doi: 10.3390/ijms25179418

Figure Lengend Snippet: Effect of GF2 on apoptosis in HeLa and SiHa cells. ( A ) Effect of GF2 on PARP and caspase 3 in HeLa and SiHa cells. ( B ) Effect of GF2 on sub-G1 phase in HeLa and SiHa cells.

Article Snippet: HeLa (ccl-2, ATCC, Manassas, VA, USA) and SiHa (HTB-35, ATCC, Manassas, VA, USA) cervical cancer cells supplied by the ATCC company were cultured in Dulbecco’s modified Eagle medium (DMEM), including 10% FBS and 1% antibiotic antimycotic solution (Welgene, Republic of Korea).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Anti-Warburg Mechanism of Ginsenoside F2 in Human Cervical Cancer Cells via Activation of miR193a-5p and Inhibition of β-Catenin/c-Myc/Hexokinase 2 Signaling Axis

doi: 10.3390/ijms25179418

Figure Lengend Snippet: Effect of GF2 on band level of Wnt, β-catenin, c-Myc, HK2, PKM2, and LDHA in HeLa and SiHa cells. ( A ) Effect of GF2 on Wnt, β-catenin, and c-Myc in HeLa and SiHa cells. ( B ) Effect of GF2 on HK2, PKM2, and LDHA in HeLa and SiHa cells. ( C ) Effect of GF2 on glucose production in HeLa and SiHa cells using a colorimetric assay. The results represent the ±SD; *** p < 0.001. ( D ) Effect of GF2 on lactate production in HeLa and SiHa cells using a colorimetric assay. The results represent mean ± SD; ** p < 0.01, *** p < 0.001.

Article Snippet: HeLa (ccl-2, ATCC, Manassas, VA, USA) and SiHa (HTB-35, ATCC, Manassas, VA, USA) cervical cancer cells supplied by the ATCC company were cultured in Dulbecco’s modified Eagle medium (DMEM), including 10% FBS and 1% antibiotic antimycotic solution (Welgene, Republic of Korea).

Techniques: Colorimetric Assay

Journal: Antioxidants

Article Title: Lindera obtusiloba Attenuates Oxidative Stress and Airway Inflammation in a Murine Model of Ovalbumin-Challenged Asthma

doi: 10.3390/antiox9070563

Figure Lengend Snippet: Effects of LOL on inflammatory cell number, Th2 cytokines, eotaxin, MUC5AC, total IgE and OVA-specific IgE in OVA-challenged asthma model. ( A ) The inflammatory cells were attached on slide and stained with Diff-Quik stain reagent. ( B ) The levels of IL-4, IL-5, IL-13, eotaxin, and MUC5AC in the BALF were determined by ELISA. ( C ) The total IgE and OVA-specific IgE in the serum were determined using ELISA. NC: normal control mice; OVA: OVA-challenged mice; DEX: dexamethasone (3 mg/kg) + OVA-challenged mice; LOL: LOL (50 or 100 mg/kg) + OVA-challenged mice. The values are expressed as the means ± SD ( n = 7/group). # p < 0.01, significantly different from NC group; †† p < 0.01, significantly different from OVA-challenged group.

Article Snippet: The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-4, IL-5, IL-6, IL-13, and eotaxin (R&D system, Minneapolis, MN, USA), MUC5AC (Cusabio Biotech Co. Wuhan, China) and IgE (BioLegend, CA, USA) were used according to the manufacturer’s instructions.

Techniques: Staining, Diff-Quik, Enzyme-linked Immunosorbent Assay, Control

Journal: Antioxidants

Article Title: Lindera obtusiloba Attenuates Oxidative Stress and Airway Inflammation in a Murine Model of Ovalbumin-Challenged Asthma

doi: 10.3390/antiox9070563

Figure Lengend Snippet: Effects of LOL on activation of MAPKs/AP-1, p65NF-κB and expression of MUC5AC in OVA-challenged asthma model. The protein levels of ( A – E ) MAPKs/AP-1, ( F ) p65NF-κB and ( G ) MUC5AC in the lung tissues were determined by western blot analysis. β-actin was used to confirm equal protein loading. NC: normal control mice; OVA: OVA-challenged asthma mice; DEX: dexamethasone (3 mg/kg) + OVA-challenged asthma mice; LOL: LOL (50 or 100 mg/kg) + OVA-challenged asthma mice. The values are expressed as the means ± SD ( n = 7/group). # p < 0.01, significantly different from NC group; †† p < 0.01, significantly different from OVA-challenged group.

Article Snippet: The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-4, IL-5, IL-6, IL-13, and eotaxin (R&D system, Minneapolis, MN, USA), MUC5AC (Cusabio Biotech Co. Wuhan, China) and IgE (BioLegend, CA, USA) were used according to the manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: Journal of leukocyte biology

Article Title: Modeling asthma: pitfalls, promises, and the road ahead

doi: 10.1002/JLB.3MR1117-436R

Figure Lengend Snippet: Shown here are eosinophils from A. Human (Homo sapiens sapiens) B. Mouse (Mus musculus) C. Horse (Equus caballus), D. Cat (Feline catus) and E. Guinea pig (Cavia porcellus). Eosinophils from human, mouse, cat and horse are cytospins preparations stained with modified Giemsa (Diff Quik) and photographed on a Leica DMI4000 microscope at original magnification of 40x. The eosinophil from guinea pig was stained with hematoxylin and eosin and imaged with a Nikon E400 microscope, Plan Fluor 100X oil objective, 1.3 N.A.

Article Snippet: The eosinophil from guinea pig was stained with hematoxylin and eosin and imaged with a Nikon E400 microscope, Plan Fluor 100X oil objective, 1.3 N.A.

Techniques: Staining, Modification, Diff-Quik, Microscopy