tbx4 lme cre  (Jackson Laboratory)

Journal: STAR Protocols

Article Title: Protocol for minicircle production for gene therapy without subsequent cleanup steps

doi: 10.1016/j.xpro.2025.103982

Figure Lengend Snippet: Gel electrophoresis of induction of mCherry-P2A- TBX4 minicircles with corresponding nanodrops Gel showing parental plasmid and two separate elutions of minicircle post-induction of mCherry-P2A-TBX4 showing clean productions. (A) shows gel electrophoresis of DNA ladder (Ladder), Parental Plasmid (PP) from preinduction miniprep and two separate elutions of minicircle (MC) from the GeneJet Maxiprep which correspond to minicircle 1 (MC1) and minicircle 2 (MC2). The minicircles are lower in size which correspond to successful induction and lack parental plasmid band noted demonstrating lack of contamination. (B) shows nanodrop quantifications from the two minicircle elutions. The minicircle elution corresponds the number on the nanodrop.

Article Snippet: TaqMan mouse Tbx4 probes , Thermo Scientific , Mm01299756_g1.

Techniques: Nucleic Acid Electrophoresis, Plasmid Preparation

Journal: STAR Protocols

Article Title: Protocol for minicircle production for gene therapy without subsequent cleanup steps

doi: 10.1016/j.xpro.2025.103982

Figure Lengend Snippet: qPCR results comparing mouse and human expression of TBX4 in NIH/3T3 cells NIH/3T3 cells were transfected with either Empty or mCherry-P2A-TBX4 minicircles and incubated for 72 h before RNA extraction and reverse transcription. Mouse and human TBX4 was analyzed in both Empty and mCherry-P2A-TBX4 minicircles compared to mouse beta-actin expression. Values above the line are p-values between compared conditions. Data are represented as mean ± SD. p -values were calculated using a two-tailed T-test. p < 0.05 was deemed significant.

Article Snippet: TaqMan mouse Tbx4 probes , Thermo Scientific , Mm01299756_g1.

Techniques: Expressing, Transfection, Incubation, RNA Extraction, Reverse Transcription, Two Tailed Test

Journal: STAR Protocols

Article Title: Protocol for minicircle production for gene therapy without subsequent cleanup steps

doi: 10.1016/j.xpro.2025.103982

Figure Lengend Snippet: Gel electrophoresis of induction of mCherry-P2A- TBX4 minicircles with corresponding nanodrops Gel showing parental plasmid and two separate elutions of minicircle post-induction of mCherry-P2A-TBX4 showing clean productions. (A) shows gel electrophoresis of DNA ladder (Ladder), Parental Plasmid (PP) from preinduction miniprep and two separate elutions of minicircle (MC) from the GeneJet Maxiprep which correspond to minicircle 1 (MC1) and minicircle 2 (MC2). The minicircles are lower in size which correspond to successful induction and lack parental plasmid band noted demonstrating lack of contamination. (B) shows nanodrop quantifications from the two minicircle elutions. The minicircle elution corresponds the number on the nanodrop.

Article Snippet: TaqMan human TBX4 probes , Thermo Scientific , Hs01057581_m1.

Techniques: Nucleic Acid Electrophoresis, Plasmid Preparation

Journal: STAR Protocols

Article Title: Protocol for minicircle production for gene therapy without subsequent cleanup steps

doi: 10.1016/j.xpro.2025.103982

Figure Lengend Snippet: qPCR results comparing mouse and human expression of TBX4 in NIH/3T3 cells NIH/3T3 cells were transfected with either Empty or mCherry-P2A-TBX4 minicircles and incubated for 72 h before RNA extraction and reverse transcription. Mouse and human TBX4 was analyzed in both Empty and mCherry-P2A-TBX4 minicircles compared to mouse beta-actin expression. Values above the line are p-values between compared conditions. Data are represented as mean ± SD. p -values were calculated using a two-tailed T-test. p < 0.05 was deemed significant.

Article Snippet: TaqMan human TBX4 probes , Thermo Scientific , Hs01057581_m1.

Techniques: Expressing, Transfection, Incubation, RNA Extraction, Reverse Transcription, Two Tailed Test

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Loss of Tbx4 Affects Postnatal Lung Development and Predisposes to Pulmonary Hypertension

doi: 10.1165/rcmb.2024-0459OC

Figure Lengend Snippet: Effective disruption of T-box 4 ( Tbx4 ) expression within the lung mesenchyme of mouse embryos. ( A ) Schematic showing the mouse Tbx4 locus and the genetics of the mice used in this study. The LME (blue box) enhancer sequence within the Tbx4 locus was used in the Tbx4 LME -Cre construct to control expression of Cre-recombinase within the developing mouse lung. The Tbx4 cond allele contains loxP sites (red arrowheads) flanking exon 5. The green boxes represent the exons, the black boxes represent the 5′ and 3′ untranslated regions, and the black arrow is the transcription start site. The control mice in the study are of the genotype Tbx4 fl/fl , and the Tbx4 conditional knockout ( Tbx4- CKO) mice are of the genotype Tbx4 LME -Cre ; Tbx4 fl/fl . ( B ) Whole-mount in situ hybridization (WMISH) using Cre antisense riboprobes to validate expression of the Cre-recombinase. Black arrow points at robust staining in the lung of an Embryonic Day (E)10.5 embryo. ( C ) WMISH using Cre antisense riboprobe on E11.5 lungs. ( D ) RNA ISH for Cre using lung tissue sections from E13.5 mouse embryos. ( E ) RNA ISH for Tbx4 using lung tissue sections from E13.5 mouse embryos. ( F–H ) RT-qPCR results using sets of primers to amplify different exons of the Tbx4 transcript. Scale bars, 100 μm. The statistical test used was a parametric unpaired two-tailed t test. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Briefly, Tbx4 LME -Cre (IMSR_JAX:033331) and Tbx4 cond (MMRRC_043812-JAX) mouse models were purchased from The Jackson Laboratory ( 16 , 17 ) and crossed to generate homozygous conditional knockouts ( Tbx4 -CKOs).

Techniques: Disruption, Expressing, Sequencing, Construct, Control, Knock-Out, In Situ Hybridization, Staining, Quantitative RT-PCR, Two Tailed Test

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Loss of Tbx4 Affects Postnatal Lung Development and Predisposes to Pulmonary Hypertension

doi: 10.1165/rcmb.2024-0459OC

Figure Lengend Snippet: Disruption of Tbx4 expression in the embryonic lung disrupts alveolar lung development. ( A ) Photomicrographs of hematoxylin and eosin–stained lung sections from mice of the indicated postnatal day. The top and bottom rows show the photos from the control and Tbx4- CKO lungs, respectively. Scale bars, 100 μm. ( B ) Mean linear intercept (MLI) calculations from lungs of the indicated time points. The graphs show individual values and mean ± SEM. Each dot represents the average MLI for a single mouse. The number of mice per group is as follows: n = 6 (Postnatal Day [P]14); n = 6 (P36); n = 10 (P180). The statistical test used was a parametric unpaired two-tailed t test. ** P < 0.01 and **** P < 0.0001.

Article Snippet: Briefly, Tbx4 LME -Cre (IMSR_JAX:033331) and Tbx4 cond (MMRRC_043812-JAX) mouse models were purchased from The Jackson Laboratory ( 16 , 17 ) and crossed to generate homozygous conditional knockouts ( Tbx4 -CKOs).

Techniques: Disruption, Expressing, Staining, Control, Two Tailed Test

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Loss of Tbx4 Affects Postnatal Lung Development and Predisposes to Pulmonary Hypertension

doi: 10.1165/rcmb.2024-0459OC

Figure Lengend Snippet: Six-month-old Tbx4- CKO lungs show pulmonary vascular simplification. ( A ) Representative photomicrographs of lung sections labeled with a von Willebrand factor antibody using immunohistochemistry. Boxes in the top right corner of the photos show a higher magnification of representative pulmonary arteries included in the analyses of this study. Scale bars, 100 μm. ( B ) Quantification of the number of von Willebrand factor–positive vessels per high-power field. The graph shows individual values and mean ± SEM. Each dot represents the average number of stained vessels per mouse lung. The statistical test used was a parametric unpaired two-tailed t test. n = 10 per group; **** P < 0.0001.

Article Snippet: Briefly, Tbx4 LME -Cre (IMSR_JAX:033331) and Tbx4 cond (MMRRC_043812-JAX) mouse models were purchased from The Jackson Laboratory ( 16 , 17 ) and crossed to generate homozygous conditional knockouts ( Tbx4 -CKOs).

Techniques: Labeling, Immunohistochemistry, Staining, Two Tailed Test

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Loss of Tbx4 Affects Postnatal Lung Development and Predisposes to Pulmonary Hypertension

doi: 10.1165/rcmb.2024-0459OC

Figure Lengend Snippet: Physiological and histological features of pulmonary hypertension in 6-month-old Tbx4- CKO mice. ( A ) Right ventricular systolic pressure (RVSP) measurements from 6-month-old mice housed in room air. We evaluated differences in RVSP in a dataset with male and female values combined, but sex-specific differences were also assessed. The graphs show individual values and mean ± SEM. Each dot represents the RVSP for a single mouse. The number of mice per group is as follows: n = 20 ( Tbx4 fl/fl ; M and F), n = 23 ( Tbx4- CKO; M and F), n = 8 ( Tbx4 fl/fl ; M), n = 10 ( Tbx4- CKO; M), n = 12 ( Tbx4 fl/fl ; F), n = 13 ( Tbx4- CKO; F). ( B ) Representative photomicrographs of pulmonary arteries from 6-month-old lung sections stained with Masson’s trichrome and Verhoeff’s stain. Scale bars, 10 μm. ( C ) The ratio of vessel wall thickness to total area of the vessel was calculated to determine the degree of vascular remodeling. The graph shows individual values and mean ± SEM. Each dot represents the calculated ratio for a single mouse. n = 10 mice per group. ( D ) The Fulton index was calculated to evaluate for right ventricular hypertrophy. The graph shows individual values and mean ± SEM. Each dot represents the Fulton index for a single mouse. n = 20 ( Tbx4 fl/fl ) and n = 22 ( Tbx4- CKO). The statistical test used was a parametric unpaired two-tailed t test. * P < 0.05, ** P < 0.01, and **** P < 0.0001.

Article Snippet: Briefly, Tbx4 LME -Cre (IMSR_JAX:033331) and Tbx4 cond (MMRRC_043812-JAX) mouse models were purchased from The Jackson Laboratory ( 16 , 17 ) and crossed to generate homozygous conditional knockouts ( Tbx4 -CKOs).

Techniques: Staining, Two Tailed Test

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Loss of Tbx4 Affects Postnatal Lung Development and Predisposes to Pulmonary Hypertension

doi: 10.1165/rcmb.2024-0459OC

Figure Lengend Snippet: RNA-sequencing analysis using P14 and P36 Tbx4- CKO lungs. ( A and B ) Heatmaps of log 2 counts per million values for relevant differentially expressed genes in Tbx4 fl/fl versus Tbx4- CKO P14 ( A ) and P36 ( B ) lungs generated using Heatmapper. The z -score for each gene across the samples was calculated to compare expression levels. A positive (red) z -score indicates higher expression of the gene, whereas a negative (blue) value indicates lower expression. A z -score close to zero indicates that the expression level for the gene is close to the mean expression level across all samples. Hierarchical clustering was used to organize and visualize relationships between the genes, and a dendrogram is shown for the sample clustering. ( C and D ) Bar graphs showing the top 10 affected regulatory pathways in P14 ( C ) and P36 ( D ) lungs, as determined by pathway analysis using MetaCore data. The x -axis shows the −log P value as calculated by MetaCore. The higher this number, the more statistically significant the pathway.

Article Snippet: Briefly, Tbx4 LME -Cre (IMSR_JAX:033331) and Tbx4 cond (MMRRC_043812-JAX) mouse models were purchased from The Jackson Laboratory ( 16 , 17 ) and crossed to generate homozygous conditional knockouts ( Tbx4 -CKOs).

Techniques: RNA Sequencing, Generated, Expressing

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: TBX4 is primarily expressed in myofibroblasts and pericytes. (A) TBX4 expression levels from a single cell RNA‐seq atlas. Dot color is indicative of expression level; size is percent of cells expressing TBX4 within that category; labels are different cell types, including five subcategories of fibroblast. (B) RT‐PCR results demonstrate that TBX4 is mainly expressed in mesenchymal origin cell lines—highest in early human lung fibroblasts (HLF) and pericytes (HLPC), lower in smooth muscle (PASMC), lowest in endothelial cells (PMVEC). (C) RNA‐Scope shows location of TBX4 RNA (red dots) and the pericyte marker Cox4i2 (white dots) with DAPI nuclear stain (blue) at postnatal Day 7. Red arrows are to draw attention to an epithelial ring (negative); yellow arrow indicates a blood vessel, likely primarily endothelium (also negative). At P7, much of the alveolar space is still mesenchyme, which we would expect to be positive. RT‐PCR, real‐time polymerase chain reaction.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: Expressing, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, RNAscope, Marker, Staining, Real-time Polymerase Chain Reaction

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: RNA‐seq outcomes. (A) Principal components analysis. HLF and HLPC are well separated, and control and TBX4 knockdown are separated; the direction of separation is distinct in HLF and HLPC, indicating different effects in the two cell types. (B) Overrepresented ontology groups in 552 genes altered at p < 0.05 and more than 1.2x in both HLF and HLPC. (C) Significantly altered matrix genes ( p < 0.05) have both discordant and concordant change with TBX4 knockout between cell types. HLF and HLPC are each normalized to their own wild‐type control; HLF are indicated by blue points and arrows, HLPC by red. HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: RNA Sequencing, Control, Knockdown, Knock-Out

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: (A) The workflow strategy for combining results from RNA‐seq and TBX4 ChIP‐seq carried out in HLF and HLPC from control and TBX4KD. There are 555 genes with altered expression in both cell types, and which contain putative TBX4 binding sites by ChIP‐seq. Circle size is proportional to number of genes; colors are arbitrary. (B) Plot of false discovery rate and enrichment ratio for overrepresented gene ontology groups among the 555 genes found in (A) above. ChIP‐seq, chromatin immunoprecipitation sequencing; HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: RNA Sequencing, ChIP-sequencing, Control, Expressing, Binding Assay

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: Ontology groups significantly overrepresented in genes bound and regulated by TBX4 in human lung fibroblasts and human lung pericyte cells.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: Activity Assay, Transduction, Sublimation

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: (A) The workflow of ChIP‐seq and following data analysis (B, C) Pie charts show the distribution of TBX4 binding sites in fibroblasts (B) and in pericytes (C) according to peak location across different genomic regions in the human genome. ChIP‐seq, chromatin immunoprecipitation sequencing; HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: ChIP-sequencing, Binding Assay

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: Migration and BrdU proliferation assays by knocking down TBX4 in HLF. (A) Time‐lapse microscopy images of migration of knockdown (KD) control (left panels) and TBX4 KD (right panels) in HLF at 0, 24 h after scratch. The dotted lines define the area lacking cells. (B) Quantification of the migration distance during 24 h. (C) Quantitative analysis of BrdU‐positive HLF TBX4 KD control and TBX4 KD cells after incubated with BrdU 48 h. BrdU, bromodeoxyuridine; HLF, human lung fibroblasts.

Article Snippet: The TBX4 antibody for Western Blot was sc‐398903 (F‐12) (Santa Cruz Biotechnology, Dallas, TX USA).

Techniques: Migration, Time-lapse Microscopy, Knockdown, Control, Incubation

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: TBX4 is primarily expressed in myofibroblasts and pericytes. (A) TBX4 expression levels from a single cell RNA‐seq atlas. Dot color is indicative of expression level; size is percent of cells expressing TBX4 within that category; labels are different cell types, including five subcategories of fibroblast. (B) RT‐PCR results demonstrate that TBX4 is mainly expressed in mesenchymal origin cell lines—highest in early human lung fibroblasts (HLF) and pericytes (HLPC), lower in smooth muscle (PASMC), lowest in endothelial cells (PMVEC). (C) RNA‐Scope shows location of TBX4 RNA (red dots) and the pericyte marker Cox4i2 (white dots) with DAPI nuclear stain (blue) at postnatal Day 7. Red arrows are to draw attention to an epithelial ring (negative); yellow arrow indicates a blood vessel, likely primarily endothelium (also negative). At P7, much of the alveolar space is still mesenchyme, which we would expect to be positive. RT‐PCR, real‐time polymerase chain reaction.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: Expressing, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, RNAscope, Marker, Staining, Real-time Polymerase Chain Reaction

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: RNA‐seq outcomes. (A) Principal components analysis. HLF and HLPC are well separated, and control and TBX4 knockdown are separated; the direction of separation is distinct in HLF and HLPC, indicating different effects in the two cell types. (B) Overrepresented ontology groups in 552 genes altered at p < 0.05 and more than 1.2x in both HLF and HLPC. (C) Significantly altered matrix genes ( p < 0.05) have both discordant and concordant change with TBX4 knockout between cell types. HLF and HLPC are each normalized to their own wild‐type control; HLF are indicated by blue points and arrows, HLPC by red. HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: RNA Sequencing, Control, Knockdown, Knock-Out

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: (A) The workflow strategy for combining results from RNA‐seq and TBX4 ChIP‐seq carried out in HLF and HLPC from control and TBX4KD. There are 555 genes with altered expression in both cell types, and which contain putative TBX4 binding sites by ChIP‐seq. Circle size is proportional to number of genes; colors are arbitrary. (B) Plot of false discovery rate and enrichment ratio for overrepresented gene ontology groups among the 555 genes found in (A) above. ChIP‐seq, chromatin immunoprecipitation sequencing; HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: RNA Sequencing, ChIP-sequencing, Control, Expressing, Binding Assay

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: Ontology groups significantly overrepresented in genes bound and regulated by TBX4 in human lung fibroblasts and human lung pericyte cells.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: Activity Assay, Transduction, Sublimation

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: (A) The workflow of ChIP‐seq and following data analysis (B, C) Pie charts show the distribution of TBX4 binding sites in fibroblasts (B) and in pericytes (C) according to peak location across different genomic regions in the human genome. ChIP‐seq, chromatin immunoprecipitation sequencing; HLF, human lung fibroblasts; HLPC, human lung pericytes.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: ChIP-sequencing, Binding Assay

Journal: Pulmonary Circulation

Article Title: RNA‐Seq and ChIP‐Seq Identification of Unique and Overlapping Target Genes and Pathways Regulated by TBX4 in Human Pulmonary Fibroblasts and Pericytes

doi: 10.1002/pul2.70058

Figure Lengend Snippet: Migration and BrdU proliferation assays by knocking down TBX4 in HLF. (A) Time‐lapse microscopy images of migration of knockdown (KD) control (left panels) and TBX4 KD (right panels) in HLF at 0, 24 h after scratch. The dotted lines define the area lacking cells. (B) Quantification of the migration distance during 24 h. (C) Quantitative analysis of BrdU‐positive HLF TBX4 KD control and TBX4 KD cells after incubated with BrdU 48 h. BrdU, bromodeoxyuridine; HLF, human lung fibroblasts.

Article Snippet: For TBX4 knockdown (KD) in HLF by TBX4 siRNA, cells were plated at a density of 1.8 × 10 6 cells/plate in a 10 cm plate, and then transfected with 150 pmol TBX4 siRNA or control siRNA by Lipofectamine RNAiMAX reagent (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions.

Techniques: Migration, Time-lapse Microscopy, Knockdown, Control, Incubation