Journal: medRxiv

Article Title: Anomalous epithelial variations and ectopic inflammatory response in chronic obstructive pulmonary disease

doi: 10.1101/2020.12.03.20242412

Figure Lengend Snippet: A. Representative images immunostained with CCL2, TNC, and CXCL8 with AT2 cell markers (SFTPC or ABCA3) in COPD and never-smoker lung sections. B . Violin plots for COL4A1, COL4A2, and COL4A3 in AT1 clusters. C. Violin plots for FKBP5 in selected cell types. a) present dataset; b) combined publicly-available datasets.

Article Snippet: Antibodies and reagents were as follows: anti-PD-L1 (rabbit, 1:1000, #ab205921, Abcam, Cambridge, UK); anti-TNC (rabbit, 1:100, #HPA004823, Sigma-Aldrich); anti-CCL2 (rabbit, 1:100, #HPA019163, Sigma-Aldrich); anti-RAGE (rabbit, 1:1000, #ab216329, Abcam); anti-SFTPC (rabbit, 1:1000, #HPA010928, Sigma-Aldrich); anti-ABCA3 (mouse, 1:1000, #WMAB-ABCA3-17, Seven Hills Bioreagents, OH, USA); anti- macrophage inflammatory protein 3 alpha (rabbit, 1:1000, #ab224188, Abcam); anti- CXCL1 (mouse, 1:100, #MAB275, R&D Systems, MN, USA); anti-CXCL8 (mouse, 1:100, #MAB208, R&D Systems); and Hoechst 33242 (1:1000, #H342, Sigma-Aldrich).

Techniques:

Journal: medRxiv

Article Title: Anomalous epithelial variations and ectopic inflammatory response in chronic obstructive pulmonary disease

doi: 10.1101/2020.12.03.20242412

Figure Lengend Snippet: A. 100% stacked bar charts of the percentage of cell populations for AT1 subtype clusters, AT2 subtype clusters, and basal lineage clusters. B. Bar charts displaying the percentages of subpopulations such as AT1-B, AT2-A, AT2-B, club, goblet, and basal clusters across patient states. C. a UMAP plot of epithelial cells focusing on the AT2-C cluster (left). The cell population of the AT2-C cluster (right). The y-axis represents the ratio of cells in the AT2-C cluster across patient states. Brackets () represent the percentage of cells of the AT2-C cluster based on epithelial cells in each of the patient states. D. Violin plots of representative inflammatory-related genes displaying the AT2-C (iAT2) cluster. E. Violin plots for CXCL1 and CXCL8 in AT2 cells in combined publicly-available datasets. F. Representative images immunostained with CD274 (PD-L1), CXCL1 and CXCL20 with AT2 cell markers (SFTPC or ABCA3) in COPD and never-smoker lung sections. Scale bars, 25 μm (left), 12.5 μm (right).

Article Snippet: Antibodies and reagents were as follows: anti-PD-L1 (rabbit, 1:1000, #ab205921, Abcam, Cambridge, UK); anti-TNC (rabbit, 1:100, #HPA004823, Sigma-Aldrich); anti-CCL2 (rabbit, 1:100, #HPA019163, Sigma-Aldrich); anti-RAGE (rabbit, 1:1000, #ab216329, Abcam); anti-SFTPC (rabbit, 1:1000, #HPA010928, Sigma-Aldrich); anti-ABCA3 (mouse, 1:1000, #WMAB-ABCA3-17, Seven Hills Bioreagents, OH, USA); anti- macrophage inflammatory protein 3 alpha (rabbit, 1:1000, #ab224188, Abcam); anti- CXCL1 (mouse, 1:100, #MAB275, R&D Systems, MN, USA); anti-CXCL8 (mouse, 1:100, #MAB208, R&D Systems); and Hoechst 33242 (1:1000, #H342, Sigma-Aldrich).

Techniques:

Journal: Veterinary Research

Article Title: Cytokine profiles in pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

doi: 10.1186/s13567-014-0113-8

Figure Lengend Snippet: Standards, capture and detection antibodies used by FMIA and ELISA

Article Snippet: IL8 (27) , SD061 (L) , MCA1660 (C) , MAB5351 (RD).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: BMC Genomics

Article Title: Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

doi: 10.1186/s12864-016-2565-x

Figure Lengend Snippet: IL8 protein production from Holstein and Angus fibroblasts. a IL8 production by dermal fibroblasts isolated from 19-month old Holstein ( n = 5) and Angus ( n = 12) heifers and challenged for 24 h with LPS (100ng/ml) or IL1B (1ng/ml). b Eight cultures ( n = 4/breed) were randomly chosen for RNA-seq and MIRA-seq analysis and levels of IL8 protein following a second 24 h challenge with either LPS (100ng/ml) or IL1B (1ng/ml) on these 8 cultures are shown. There was no detectable IL8 production from un-treated cultures of both breeds. Values are mean ± SEM. * indicates P < 0.05, ** indicates P < 0.01

Article Snippet: Mouse anti-bovine (clone 170.13, gifted by Samuel Maheswaren, University of Minnesota, St. Paul, MN) and a biotinylated goat anti-human IL8 (R&D Systems Inc., Minneapolis, MN) were used as capture and detection antibodies, respectively.

Techniques: Isolation, RNA Sequencing

Journal: BMC Genomics

Article Title: Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

doi: 10.1186/s12864-016-2565-x

Figure Lengend Snippet: Average response to LPS from combined Holstein and Angus cultures measured by increases in expression of immune-associated genes compared to hour 0 post-LPS

Article Snippet: Mouse anti-bovine (clone 170.13, gifted by Samuel Maheswaren, University of Minnesota, St. Paul, MN) and a biotinylated goat anti-human IL8 (R&D Systems Inc., Minneapolis, MN) were used as capture and detection antibodies, respectively.

Techniques: Expressing, Activation Assay, Ubiquitin Proteomics, Virus

Journal: BMC Genomics

Article Title: Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

doi: 10.1186/s12864-016-2565-x

Figure Lengend Snippet: Differential gene expression (fold change) of immune-associated genes at hours 0, 2, and 8 post-LPS treatment of fibroblasts collected from Holstein animals as compared to fibroblasts collected from Angus animals

Article Snippet: Mouse anti-bovine (clone 170.13, gifted by Samuel Maheswaren, University of Minnesota, St. Paul, MN) and a biotinylated goat anti-human IL8 (R&D Systems Inc., Minneapolis, MN) were used as capture and detection antibodies, respectively.

Techniques: Gene Expression, DNA Methylation Assay

Journal: BMC Genomics

Article Title: Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

doi: 10.1186/s12864-016-2565-x

Figure Lengend Snippet: Selected gene expression by Holstein and Angus fibroblasts in response to LPS. Differences in expression of TLR4 ( a ) IL8 ( b ) TNF ( c ) CCL20 ( d ) and CCL5 ( e ) between Holstein and Angus fibroblast cultures at hours 0, 2, and 8 post-LPS exposure were determined by RT-qPCR. Values are expressed as delta Ct, or the difference in cycles to threshold (Ct) between the gene of interest and the endogenous gene control beta-actin ( ACTB) . Fold differences (2 -ΔΔCt ) in expression between Holstein and Angus cultures are indicated in parentheses. All values are mean ± SEM. ( n = 4/group). * P < 0.05 and ** P < 0.01 ( t -test)

Article Snippet: Mouse anti-bovine (clone 170.13, gifted by Samuel Maheswaren, University of Minnesota, St. Paul, MN) and a biotinylated goat anti-human IL8 (R&D Systems Inc., Minneapolis, MN) were used as capture and detection antibodies, respectively.

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Control

Journal: BMC Genomics

Article Title: Differential responsiveness of Holstein and Angus dermal fibroblasts to LPS challenge occurs without major differences in the methylome

doi: 10.1186/s12864-016-2565-x

Figure Lengend Snippet: RT-qPCR primer pairs used for amplification of target genes

Article Snippet: Mouse anti-bovine (clone 170.13, gifted by Samuel Maheswaren, University of Minnesota, St. Paul, MN) and a biotinylated goat anti-human IL8 (R&D Systems Inc., Minneapolis, MN) were used as capture and detection antibodies, respectively.

Techniques: Amplification, Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: The treatment with IL6 and IL8 induced morphological and phenotypic changes associated with an EMT process. a MCF-7 cells stimulated with 50 ng/ml of IL6, IL8 or IL6 + IL8 cytokines. 5 ng/ml of TGF-β was used as a positive control and MCF-7 cultured in MEM-α medium supplemented with 0,5% FBS was used as a negative control. The evaluation was done after 96 h of cytokine treatment. Representative images are shown. Arrowhead indicates cells with fibroblastoid morphology. Scale bar, 10 μm. b Representative plot of fluorescence intensity of the EMT surface markers was determined by FACS using a CD44-FITC and a CD24-PE monoclonal antibody in MCF-7 cells stimulated for 5 days with cytokines ( n = 2)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Positive Control, Cell Culture, Negative Control, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: The treatment with IL6 and IL8 increases the migration capacity of MCF-7 cells. a Wound healing assays performed in MCF-7 cells treated with cytokines. Representative micrographs (10×) were taken at 0, 6 and 12 h showing an increased migration capacity of treated MCF-7 cells. Black dotted line indicated the area of the wound. b Quantification of the area of wound healing assay from at least 10 images by using the image J program. Error bars indicate SEM. (** p < 0.01). c Transwell migration assay towards 20% FBS performed in cytokine-treated and control MCF-7 cells. Representative micrographs (20×) were taken from the membrane filter (bottom surface of filters) stained with Crystal violet. d Quantification of transwell migration assay by counting the number of cells present in the lower compartment. Error bars indicate SEM. (* p < 0.05). e Transwell matrigel invasion assay towards 20% FBS performed in cytokine-treated and control MCF-7 cells. The histograms show the number of cells present on the bottom surface of filters from at least 15 images. Error bars indicate SEM. (*** p < 0.001) ( n = 2)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Migration, Wound Healing Assay, Transwell Migration Assay, Control, Membrane, Staining, Invasion Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: The treatment with IL6 and IL8 induced gene expression and functional changes associated with an EMT program. Gene expression levels of EMT-associated transcription factors Zeb1, Twist 1 and Twist 2 ( a ) and Snail 1 and Snail2/Slug ( b ) in MCF-7 cells stimulated with cytokines as indicated and determined by qRT–PCR. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM. (* p < 0.05; ** p < 0.01; *** p < 0.001) ( n = 2). c Adhesion of MCF-7 cells to immobilized-fibronectin after cytokine treatment. Representative micrographs (20×) taken from the plates stained with Crystal violet. The number of adherent cells was counted. Error bars indicate SEM. (* p < 0.05; ** p < 0.01)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Gene Expression, Functional Assay, Quantitative RT-PCR, Control, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: MCF-7 cells treated with IL6 and IL8 exhibit self-renewal properties. a Sphere formation assay in the presence of defined medium (EGF and FGF) in MCF-7 cells that were previously treated or not (Control) with SCM ( a and b ) or with cytokines ( c and d ) during 5 days. Representative micrographs (10×) show sphere formation after 8 days of induction. Scale bar, 100 μm. The total number of spheres per well larger than 100 μm was counted at day 8. Error bars indicate SEM. (** p < 0.01) ( n = 2)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Tube Formation Assay, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: MCF-7 cells treated with IL6 and IL8 exhibit differentiation potential characteristics and express reprogramming factors. a Differentiation to mesenchymal cell lineages was induced by using specific induction media as described in Materials and Methods and evaluated after 10 days of induction. Osteoblasts (ALP activity), adipocytes (oil red-O) and chondrocytes (Safranin O) differentiation was determined with specific staining. Representative images are shown. Scale bar, 10 μm. b Gene expression levels of reprogramming-associated transcription factors in MCF-7 cells stimulated with cytokines as indicated and determined by qRT–PCR. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM (* p < 0.05) ( n = 2)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Activity Assay, Staining, Gene Expression, Quantitative RT-PCR, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: The pro-inflammatory cytokines IL6 and IL8 are required for the maintenance of the migration capacity and stem cell properties in MCF-7 cells. MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies. The histograms show the quantification of the area of wound healing from at least 10 images taken at 12, 24 and 36 h, using the image J program. Error bars indicate SEM. (*** p < 0.001). b Sphere formation assay in the presence of defined medium (EGF and FGF) in MCF-7 cells that were previously treated or not (Control) with SCM alone or in the presence of neutralizing antibodies. The total number of primary, secondary and tertiary spheres per well larger than 100 μm was counted at day 5. Error bars indicate SEM. (*** p < 0.001) ( n = 2). c Representative micrographs (10×) showing primary, secondary and tertiary generations of mammospheres. Scale bar, 100 μm

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Migration, Incubation, Bioprocessing, Tube Formation Assay, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Senescence-associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line

doi: 10.1186/s12964-017-0172-3

Figure Lengend Snippet: The treatment with IL6 and IL8 induces senescence in MCF-7 cells. a Representative images of MCF-7 cells treated with SCM during 10 days or ( c ) cytokines (50 ng/ml) during 5 days and stained for SA-β-GAL. Scale bar, 10 μm. b Gene expression profile of p16, p21 and p53 in MCF-7 cells stimulated with SCM or ( e ) cytokines, as indicated. The values were normalized to GADPH and relative to control cells ( dotted lines ). Error bars represent SEM. (* p < 0.05; ** p < 0.01; *** p < 0.001) ( n = 2). The incubation of SCM with neutralizing anti-IL6 and anti-IL8 antibodies suppresses SA-βGal activity ( d ) and reduce the expression of senescent markers P16, P21 and P53 ( f ) determined by qRT-PCR. Error bars represent SEM. (** p < 0.01; *** p < 0.001) ( n = 2)

Article Snippet: MCF-7 cells were incubated with SCM with or without 1 μg/ml of neutralizing monoclonal antibodies against IL6 (Ref: MAB206, R&D systems) and IL8 (Ref: AF-208-NA, R&D systems). a Wound healing assays showing a decrease in the migration capacity of treated MCF-7 cells with the neutralizing antibodies.

Techniques: Staining, Gene Expression, Control, Incubation, Activity Assay, Expressing, Quantitative RT-PCR