Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: (a) Protocol. (b) Typical calcium (Ca 2+ ) transients before MI in C57BL/6J mice in the “remote zone” (“RZ”) and “border zone” (“BZ”). (c) Mean Ca 2+ transient peak, (d) mean Ca 2+ transient amplitude n =76 (9 hearts) and from the BZ n =43 (9 hearts), (e) Mean caffeine-induced Ca 2+ transient amplitude and (f) mean sarcoendoplasmic reticulum calcium transport ATPase (SERCA) activity from C57BL/6J mice from RZ, n =76 (8 hearts) and from the BZ n =43 (8 hearts). (g) Typical Ca 2+ transients 1 day post-MI in C57BL/6J mice in the RZ and BZ. (h) Mean Ca 2+ transient peak, (i) mean Ca 2+ transient amplitude, (j) mean caffeine-induced Ca 2+ transient amplitude and (k) mean SERCA activity from C57BL/6J 1-day post-MI mice from RZ, n =64 (7 hearts) and from the BZ n =30 (7 hearts). (l) Typical Ca 2+ transients 1-day post-MI in Runx1 Δ/Δ mice in the RZ and BZ. (m) Mean Ca 2+ transient peak, (n) mean Ca 2+ transient amplitude, (o) mean caffeine induced Ca 2+ transient amplitude and (p) mean SERCA activity from Runx1 Δ/Δ mice 1-day post-MI mice from RZ, n =16 (4 hearts) and from the BZ n =9 (4 hearts). Error bars represent mean ± SEM. ** P <0.01, *** P <0.001.

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: (a) Schematic of comparison and Venn diagram of gene differences. (b) Volcano plots of differentially regulated genes unique to Runx1 fl/fl mice (left, black) and Runx1 Δ/Δ mice (right, red) in the BZ versus the RZ at 1 day post-MI. Top five upregulated and down regulated genes based on false discovery rate (FDR) are noted. (c) Enriched biological pathways ranked by logP (B-H p value) using IPA analysis from unique differences in Runx1 fl/fl mice (circles) compared with BZ and RZ differences in Runx1 Δ/Δ mice (triangles). Blue-to-orange heatmap and symbol colours represent predicted inhibition (blue) and activation (orange) or no change (grey) of pathways based on Z-score.

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Comparison, Inhibition, Activation Assay

Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: Schematic visualisation of genes involved in oxidative phosphorylation complexes from the RNAseq data from the border zone (BZ) and remote zone (RZ) in Runx1 fl/fl and Runx1 Δ/Δ mice 1 day post-MI in Ingenuity Pathway Analysis (IPA) (downregulated – green, upregulated – red, no difference – white) of (a) Runx1 fl/fl mice (7166 differentially expression genes) and (b) Runx1 Δ/Δ mice (1748 differentially expressed genes). Low resolution electron microscopy images from BZ of (c) Runx1 fl/fl mice and (d) Runx1 Δ/Δ mice. Mitochondria are outlined in yellow, with damaged mitochondria indicated by yellow arrows. High-resolution electron tomography representative images of (e-f) damaged mitochondria from the BZ of Runx1 fl/fl mice 1 day post-MI and (g) healthy mitochondria from the BZ of Runx1 Δ/Δ mice 1 day post-MI. Quantification of (h) mitochondrial density as a percentage of cell area, (i) the number of damaged mitochondria as a percentage of the total number of mitochondria, and (j) average mitochondrial size from electron microscopy images of the BZ from Runx1 fl/fl mice and Runx1 Δ/Δ mice 1 day post-MI n =92 (2 hearts). Error bars represent mean ± SEM. * P <0.05, unpaired Student t-test on average heart data.

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Phospho-proteomics, Expressing, Electron Microscopy, Tomography

Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: ( a ) Schematic representation of Ad given via an injection into the border zone immediately after induction of MI. ( b-e ) Typical images of regional heart sections by RNA i n situ hybridisation (using RNAscope). Regions examined were the remote zone (RZ), infarct zone (IZ) and border zone (BZ) at 7 days post-MI in Ad- Runx1 -shRNA injected mice ( n =4) and Ad-scramble-shRNA injected mice ( n =4). Probes for Runx1 (pink punctate dots) and pericentriolar material 1 ( PCM-1 ) (brown punctate dots) were used. Scale bar, 10 µm; magnification 100x. ( c ) Mean quantification of Runx1- positive cardiomyocyte nuclei ( PCM-1 + and Runx1 ) expressed as a percentage of total nuclei (data from b). * P <0.05, 7 days after MI Ad- Runx1 -shRNA vs. Ad-scramble-shRNA. ( d ) Echocardiography (scale: x=0.1 s; y=2 mm). ( e ) The 7-day echocardiographic data for fractional shortening (FS) of Ad- Runx1 -shRNA ( n =8; day 0, n =8; day 1, n =7; day 2, n =7; day 7) vs. Ad-scramble-shRNA (n=8; day 0, n =8; day 1, n =7; day 2, n =7; day 7). ( f ) Typical picrosirius-red-stained hearts and infarct size as the percentage of the left ventricle ( n =3). ( g ) Schematic representation of AAV given via tail vein injection ( h ) Typical images of regional heart sections by RNA i n situ hybridisation (using RNAscope). Regions examined were the RZ (AAV- Runx1 -shRNA [ n =3] vs. AAV-scramble-shRNA injected mice [ n =3]), IZ (AAV- Runx1 -shRNA [ n =3] vs. AAV-scramble-shRNA injected mice [ n =3]), and BZ (AAV- Runx1 -shRNA [ n =6] vs. AAV-scramble-shRNA injected mice [ n =5]), at 7 days post-MI. Probes for Runx1 (pink punctate dots) and pericentriolar material 1 ( PCM-1 ) (brown punctate dots) were used. Scale bar, 10 µm; magnification 100x. ( i ) Mean quantification of Runx1 positive cardiomyocyte nuclei ( PCM-1 + and Runx1 ) expressed as a percentage of total nuclei (data from h). *= P <0.05, 7 days after MI AAV- Runx1 -shRNA vs. AAV-scramble-shRNA. ( j ) Echocardiography (scale: x=0.1 s; y=2 mm). ( k ) The 7-day echocardiographic data for fractional shortening (FS) (AAV- Runx1 -shRNA [ n =9; day 0, n =9; day 1, n =9; day 7] vs. AAV-scramble-shRNA [ n =10; day 0, n =10; day 1, n =9; day 7]). ( l ) Typical picrosirius-red stained hearts and infarct size as the percentage of the left ventricle AAV- Runx1 -shRNA injected mice ( n =8) vs. AAV-scramble-shRNA injected mice ( n =6)). Error bars represent mean ± SEM. * P <0.05, Student t test.

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Injection, Hybridization, RNAscope, shRNA, Staining

Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: (a) Protocol. ( b ) The 7-day echocardiographic data for fractional shortening (FS) for mice receiving protocol 1: Ro5-3335-treated MI mice ( n =6; day 0, n =6; day 1, n =5; day 3, n =5; day 7) vs. vehicle (DMSO)-treated MI mice ( n =6; day 0, n =5; day 1, n =3; day 3, n =4; day 7). ( c ) The 7-day echocardiographic data for FS for mice receiving protocol 2: Ro5-3335-treated mice ( n =8; day 0, n =8; day 1, n =6; day 3, n =6; day 7) vs. vehicle (DMSO)-treated mice ( n =7; day 0, n =8; day 1, n =3; day 3, n =7; day 7). * P <0.05, Student t test. ( d ) Runx1 mRNA expression relative to Peptidylprolyl Isomerase B ( Ppib) as measured by real-time quantitative polymerase chain reaction (qPCR) in LV tissue from Ro5-3335-treated MI mice ( n =6) vs. vehicle (DMSO)-treated MI mice ( n =4). *** P <0.005, Student t test. ( e ) Protocol. ( f ) Runx1 mRNA expression relative to Ppib as measured by qPCR in LV tissue from Cbfβ Δ/Δ MI mice ( n =4) vs. Cbfβ fl/fl MI mice ( n =4). ( g) Runx1 mRNA expression relative to Ppib as measured by qPCR in LV isolated cardiomyocytes from Cbfβ Δ/Δ MI mice ( n =9) vs. Cbfβ fl/fl MI mice ( n =8). * P <0.05, Student t test. ( h ) The 7-day echocardiographic data for FS for from Cbfβ Δ/Δ MI mice ( n =8; day 0, n =6; day 1, n =6; day 7) vs. Cbfβ fl/fl MI mice ( n =6; day 0, n =;5 day 1, n =6; day 7). Error bars represent mean ± SEM. * P <0.05, Student t test.

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation

Journal: bioRxiv

Article Title: Inhibiting Runx1 protects heart function after myocardial infarction

doi: 10.1101/2022.02.17.480749

Figure Lengend Snippet: (a) Schematic of comparison and Venn diagram of gene expression. (b) Volcano plots of differentially regulated genes unique to Cbfβ fl/fl mice (left, black) and Cbfβ Δ/Δ mice (right, green) 7 days post-MI compared to pre-MI. Top five upregulated and down regulated genes based on false discovery rate (FDR) noted. (c) Enriched biological pathway from ingenuity pathway analysis (IPA) analysis generated from unique differences in Runx1 fl/fl (border zone, BZ vs remote zone, RZ) and Cbfβ fl/fl (post-MI vs Pre-MI) mice ranked on Z-score. Blue to orange heatmap and symbol colours represent predicted inhibition (blue) and activation (orange) or no change (white) of pathways based on Z-score. Schematic representation of genes involved in oxidative phosphorylation complexes from RNAseq IPA pathway analysis generated from all gene differences (downregulated – green, upregulated – red, no difference – white) between day 7 post-MI and pre-MI in (d) Cbfβ fl/fl (7860 differentially expressed genes) and (e) Cbfβ Δ/Δ mice (2995 differentially expressed genes).

Article Snippet: The Ad- Runx1 -shRNA and a random scramble sequence (Ad-scramble-shRNA) were purchased from Vector Biolabs (USA; Gene ID: 12394).

Techniques: Comparison, Gene Expression, Generated, Inhibition, Activation Assay, Phospho-proteomics

Journal: Oncotarget

Article Title: Synergistic cytotoxic activity of cannabinoids from cannabis sativa against cutaneous T-cell lymphoma (CTCL) in-vitro and ex-vivo

doi: 10.18632/oncotarget.27528

Figure Lengend Snippet: PBL were isolated from blood samples of Sézary patients ( n = 6), and were treated with S4 (5 μg/mL), S5 (6 μg/mL), S4 (5 μg/mL) +S5 (6 μg/mL) and control (methanol; Vehicle treatment) for 48 h. Cells were harvested and analyzed by FACS following CD4-APC, CD26-alexa 405, Annexin V-FITC and PI staining. Apoptotic-induced cells (Annexin positive cells) were determined in the CD4 + CD26 - cell population and in non-CD4 + CD26 - cells of treated cells minus control. ( A ) The percent of apoptotic-induced CD4 + CD26 - cells is presented for single treatment compared to combined treatment; ( n = 4); *** denote significant difference between means (one way ANOVA; p < 0.001). ( B ) The percent of apoptotic-induced cells following the combined treatment was compared between CD4 + CD26 - cells and non-CD4 + CD26 - cells of SPBL; ( n = 6); ** denote significant difference between means (paired student T test; 0.001 < P < 0.05).

Article Snippet: Cells were suspended in 100 μL binding buffer with 1 μL Annexin V-FITC (4830-01-1, eBioscience) + 2 μL CD26 Alexa Fluor 405-conjugated antibody (FAB1180V-100UG, R&D SYSTEMS) + 10 μL CD4-APC-conjugated antibody (FAB3791A-100, R&D SYSTEMS) and incubated for 15 min at room temperature.

Techniques: Isolation, Control, Staining

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: Summary of confocal analysis of control E12 MLPCs, E12 AT2-like cells and SAEpis.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Control

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: Confocal analysis of undifferentiated E12 control MLPCs, differentiated E12 AT2-like cells and SAEpis. Cells were labeled with primary antibodies (TERT, EpCAM, CD26 and TM4SF1) and secondary antibodies labeled with Alexa Fluor 594. Positive cells are stained red. DAPI staining shows the nuclei as blue. Negative cells are indicated only by blue nuclei. The results are representative of at least six different culture experiments and determinations done on different days.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Control, Labeling, Staining

Journal: Cytotherapy

Article Title: Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins

doi: 10.1016/j.jcyt.2021.07.017

Figure Lengend Snippet: The qRT-PCR analysis of ACE-2, DPP4 (CD26), EpCAM, SPC, CK19 and TM4SF1 expression of control E12 MLPCs, E12 AT2-like cells and SAEpis. Data were normalized to GAPDH. The results represent at least four different culture experiments done on different days.

Article Snippet: Antibodies specific for cytokeratin 19 (CK19) (MAB3608), epithelial cell adhesion molecule (EpCAM) (MAB9601), SOX17 (MAB1924), CD26 (MAB1180) and ACE-2 labeled with Alexa Fluor 594 (FAB9332T) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Quantitative RT-PCR, Expressing, Control