Review

crispr activation system for tet1 (Santa Cruz Biotechnology)

Santa Cruz Biotechnology is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Santa Cruz Biotechnology crispr activation system for tet1

AKG-TET deficiency leads to replicative senescence aHDFs (3 × 10 5 cells/mL) were seeded in each culture plate and were treated, in triplicate, without (UT) or treated (T) with C35 (5 μM), peptides (20 μM), <t>TET1</t> siRNA ( TET1 i ; 300 nM). Cells treated with Bleomycin (Bleo; 5 μg/mL), or H 2 O 2 (100 μM) were used as positive control. Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Heatmap of TET gene expression. (B) TET activity and global DNA levels of 5 mC, 5hmC, and 5 fC. (C) Expression patterns of energy/stress and nutrient-sensing pathways in aging PBMCs mirror those observed in young, actively replicating aHDFs (week 3) undergoing senescence, as well as those treated with TET1 i , C35, and RLS. (D) Expression level of hTERT , NAMPT , and PCNA . (E) Expression level of COL1A1 and ELN. (F) Cellular UPS activity, ATP, NAD + /NADH ratio, and LC3B levels. (G) Cellular levels of LC3B without (−) and after pre-treatment with (+) Bafilomycin A1 (1 μM) for 24 h. (H) Cellular ROS. (I) Extent of oxidative DNA damage assessed by the cellular level of 8OHdG. Positive control was comprised of cells treated with H 2 O 2 and Bleomycin (Bleo). Negative control was comprised of cells treated with CLV. (J) γH2AX immunolocalized to nuclei. For positive control, cells were treated with Bleomycin (Bleo). Intranuclear γH2AX (green) appears as focal spots (red arrows) or as a diffuse pan-nuclear pattern (yellow arrows) in nuclei marked by DAPI (blue) staining. Nuclei are further highlighted by double hashed lines. The boxes in the left pane (scale bars 10 μM) are magnified in the right insets (scale bars 2.5 μm). (K) NFKB1 gene expression. (L) Level of intra-nuclear phosphorylated p65 (p65P). (M) Cellular levels of ROS, IL6, and IL8 were secreted into culture media. (N) Expression levels of senescence markers ( CDKN2A , CDKN1A , CDKN1B , LTA4H , TIMP1 , and MMP1 ). (O) Expression level of LDH toxicity, SAβ-Gal activity, and extracellular level of lactic acid (LA). (P) LDHA1 gene expression. (Q) Rate of proliferation assessed by the quantitation of BrdU incorporated into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. Points are shown as empty and mean points as filled circles. The distribution of data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Crispr Activation System For Tet1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/crispr+activation+system+for+tet1/pmc12767804-111-0-10?v=Santa+Cruz+Biotechnology
Average 90 stars, based on 2 article reviews

crispr activation system for tet1 - by Bioz Stars, 2026-07

90/100 stars

Images

1) Product Images from "AKG-TET axis is central to senescence plasticity"

Article Title: AKG-TET axis is central to senescence plasticity

Journal: iScience

doi: 10.1016/j.isci.2025.114298

Figure Legend Snippet: AKG-TET deficiency leads to replicative senescence aHDFs (3 × 10 5 cells/mL) were seeded in each culture plate and were treated, in triplicate, without (UT) or treated (T) with C35 (5 μM), peptides (20 μM), TET1 siRNA ( TET1 i ; 300 nM). Cells treated with Bleomycin (Bleo; 5 μg/mL), or H 2 O 2 (100 μM) were used as positive control. Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Heatmap of TET gene expression. (B) TET activity and global DNA levels of 5 mC, 5hmC, and 5 fC. (C) Expression patterns of energy/stress and nutrient-sensing pathways in aging PBMCs mirror those observed in young, actively replicating aHDFs (week 3) undergoing senescence, as well as those treated with TET1 i , C35, and RLS. (D) Expression level of hTERT , NAMPT , and PCNA . (E) Expression level of COL1A1 and ELN. (F) Cellular UPS activity, ATP, NAD + /NADH ratio, and LC3B levels. (G) Cellular levels of LC3B without (−) and after pre-treatment with (+) Bafilomycin A1 (1 μM) for 24 h. (H) Cellular ROS. (I) Extent of oxidative DNA damage assessed by the cellular level of 8OHdG. Positive control was comprised of cells treated with H 2 O 2 and Bleomycin (Bleo). Negative control was comprised of cells treated with CLV. (J) γH2AX immunolocalized to nuclei. For positive control, cells were treated with Bleomycin (Bleo). Intranuclear γH2AX (green) appears as focal spots (red arrows) or as a diffuse pan-nuclear pattern (yellow arrows) in nuclei marked by DAPI (blue) staining. Nuclei are further highlighted by double hashed lines. The boxes in the left pane (scale bars 10 μM) are magnified in the right insets (scale bars 2.5 μm). (K) NFKB1 gene expression. (L) Level of intra-nuclear phosphorylated p65 (p65P). (M) Cellular levels of ROS, IL6, and IL8 were secreted into culture media. (N) Expression levels of senescence markers ( CDKN2A , CDKN1A , CDKN1B , LTA4H , TIMP1 , and MMP1 ). (O) Expression level of LDH toxicity, SAβ-Gal activity, and extracellular level of lactic acid (LA). (P) LDHA1 gene expression. (Q) Rate of proliferation assessed by the quantitation of BrdU incorporated into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. Points are shown as empty and mean points as filled circles. The distribution of data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Techniques Used: Positive Control, Incubation, Gene Expression, Activity Assay, Expressing, Negative Control, Staining, Quantitation Assay

AKG-TET dependent resilience to damage and protection against damage-induced senescence Proliferating aHDFs (0.3 × 10 6 /mL) and PBMCs (70 years; PBMC 70Yr, 1 × 10 6 /mL) were seeded in culture plates and pre-treated in triplicate without (untreated: UT) or with H 2 O 2 (100 μM) for 24 h. After 24 h, cells were washed and treated without or with CLV (20 μM), or CRISPR TET1 ( TET1 CR ;1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression levels of TETs in PBMCs. (B) AKG bioavailability in PBMCs. (C) Expression levels of energy/stress and nutritional sensors in PBMCs. (D) Heatmap of senescence marker gene expression in PBMCs. (E) Level of extracellular lactic acid (LA) and SAβ-Gal activity in PBMCs. (F) Quantification of BrdU and ROS (qROS) in PBMCs. Bar and line graphs show the means ± SD. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 . Points are shown as empty and mean points as filled circles.

Figure Legend Snippet: AKG-TET dependent resilience to damage and protection against damage-induced senescence Proliferating aHDFs (0.3 × 10 6 /mL) and PBMCs (70 years; PBMC 70Yr, 1 × 10 6 /mL) were seeded in culture plates and pre-treated in triplicate without (untreated: UT) or with H 2 O 2 (100 μM) for 24 h. After 24 h, cells were washed and treated without or with CLV (20 μM), or CRISPR TET1 ( TET1 CR ;1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression levels of TETs in PBMCs. (B) AKG bioavailability in PBMCs. (C) Expression levels of energy/stress and nutritional sensors in PBMCs. (D) Heatmap of senescence marker gene expression in PBMCs. (E) Level of extracellular lactic acid (LA) and SAβ-Gal activity in PBMCs. (F) Quantification of BrdU and ROS (qROS) in PBMCs. Bar and line graphs show the means ± SD. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 . Points are shown as empty and mean points as filled circles.

Techniques Used: CRISPR, Incubation, Expressing, Marker, Gene Expression, Activity Assay

AKG-TET deficient senescence state is reversible Young replicatively proliferating aHDFs (week 3, 0.3 × 10 6 /mL)) were seeded in culture plates and pre-treated in triplicates without (UT) and with C35 (5 μM), RLS (20 μM), or TET1 siRNA ( TET1 i ; 300 nM). Aliquot of cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. The remaining cells were re-seeded in equal numbers and cultured without treatment for an additional 7 days (withdrawal). Cultures were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media. (A) Quantification of TET activity, and 5 mC and 5fc levels. (B) Expression level of energy/stress and nutritional sensor genes. (C) Expression level of NFKB1 , RELA , IKBA , COL1A1, and ELN . (D) γH2AX immunolocalized (cyan) in nuclei. Nuclei stained for DAPI (deep blue) are marked by double hashed lines. (E) Extracellular level of lactic acid (LA), SAβ-Gal activity, LDH toxicity, and UPS. (F) Intracellular level of ROS and 8OHdG. (G) Heatmap of senescence markers and RRM2 gene expression. (H) Rate of proliferation assessed by BrdU incorporation into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Figure Legend Snippet: AKG-TET deficient senescence state is reversible Young replicatively proliferating aHDFs (week 3, 0.3 × 10 6 /mL)) were seeded in culture plates and pre-treated in triplicates without (UT) and with C35 (5 μM), RLS (20 μM), or TET1 siRNA ( TET1 i ; 300 nM). Aliquot of cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. The remaining cells were re-seeded in equal numbers and cultured without treatment for an additional 7 days (withdrawal). Cultures were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media. (A) Quantification of TET activity, and 5 mC and 5fc levels. (B) Expression level of energy/stress and nutritional sensor genes. (C) Expression level of NFKB1 , RELA , IKBA , COL1A1, and ELN . (D) γH2AX immunolocalized (cyan) in nuclei. Nuclei stained for DAPI (deep blue) are marked by double hashed lines. (E) Extracellular level of lactic acid (LA), SAβ-Gal activity, LDH toxicity, and UPS. (F) Intracellular level of ROS and 8OHdG. (G) Heatmap of senescence markers and RRM2 gene expression. (H) Rate of proliferation assessed by BrdU incorporation into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Techniques Used: Incubation, Cell Culture, Activity Assay, Expressing, Staining, Gene Expression, BrdU Incorporation Assay

Activation of the AKG-TET axis reverses replicative and age induced senescence Replicatively induced senescence. Replicatively senescent aHDFs (0.3 × 10 6 cells/mL) were seeded in triplicate in each plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids (TET1 CR , 1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression level of TET s. (B) AKG bioavailability and TET activity (TET act ). (C) Expression level of energy/stress and nutritional sensor gene expression. (D) Intracellular level of ROS. (E) Heatmap of senescence marker and RRM2 gene expression. (F) Level of lactic acid (LA) in culture media. (G) Cellular SAβ-Gal activity. (H) Quantitation of nuclear BrdU. (I) Total cell numbers. Age induced senescence. PBMCs (PBMC 70Yr ) were seeded (1 × 10 6 cells/mL) in triplicate in each well of a 24-well plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids ( TET1 CR : 1 μg/mL). Cells and culture media were removed on day 7 of treatment for analysis. (J) Quantitation of bioavailable AKG and TET activity in cells treated with RLS versus CLV. (K) qPCR quantitation of the expression level of TET s. (L) qPCR quantitation of the expression level of energy/stress and nutritional sensor gene expression. (M) Quantitation of BrdU incorporated into nuclei of cells in S-phase, lactic (LA) acid in culture media, cellular SAβ-Gal activity, and ROS. (N) Heatmap of senescence markers and RRM2 in PBMCs. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 , ∗∗∗∗p ≤ 5 × 10 −5 .

Figure Legend Snippet: Activation of the AKG-TET axis reverses replicative and age induced senescence Replicatively induced senescence. Replicatively senescent aHDFs (0.3 × 10 6 cells/mL) were seeded in triplicate in each plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids (TET1 CR , 1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression level of TET s. (B) AKG bioavailability and TET activity (TET act ). (C) Expression level of energy/stress and nutritional sensor gene expression. (D) Intracellular level of ROS. (E) Heatmap of senescence marker and RRM2 gene expression. (F) Level of lactic acid (LA) in culture media. (G) Cellular SAβ-Gal activity. (H) Quantitation of nuclear BrdU. (I) Total cell numbers. Age induced senescence. PBMCs (PBMC 70Yr ) were seeded (1 × 10 6 cells/mL) in triplicate in each well of a 24-well plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids ( TET1 CR : 1 μg/mL). Cells and culture media were removed on day 7 of treatment for analysis. (J) Quantitation of bioavailable AKG and TET activity in cells treated with RLS versus CLV. (K) qPCR quantitation of the expression level of TET s. (L) qPCR quantitation of the expression level of energy/stress and nutritional sensor gene expression. (M) Quantitation of BrdU incorporated into nuclei of cells in S-phase, lactic (LA) acid in culture media, cellular SAβ-Gal activity, and ROS. (N) Heatmap of senescence markers and RRM2 in PBMCs. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 , ∗∗∗∗p ≤ 5 × 10 −5 .

Techniques Used: Activation Assay, CRISPR, Incubation, Expressing, Activity Assay, Gene Expression, Marker, Quantitation Assay

Image Search Results

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficiency leads to replicative senescence aHDFs (3 × 10 5 cells/mL) were seeded in each culture plate and were treated, in triplicate, without (UT) or treated (T) with C35 (5 μM), peptides (20 μM), TET1 siRNA ( TET1 i ; 300 nM). Cells treated with Bleomycin (Bleo; 5 μg/mL), or H 2 O 2 (100 μM) were used as positive control. Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Heatmap of TET gene expression. (B) TET activity and global DNA levels of 5 mC, 5hmC, and 5 fC. (C) Expression patterns of energy/stress and nutrient-sensing pathways in aging PBMCs mirror those observed in young, actively replicating aHDFs (week 3) undergoing senescence, as well as those treated with TET1 i , C35, and RLS. (D) Expression level of hTERT , NAMPT , and PCNA . (E) Expression level of COL1A1 and ELN. (F) Cellular UPS activity, ATP, NAD + /NADH ratio, and LC3B levels. (G) Cellular levels of LC3B without (−) and after pre-treatment with (+) Bafilomycin A1 (1 μM) for 24 h. (H) Cellular ROS. (I) Extent of oxidative DNA damage assessed by the cellular level of 8OHdG. Positive control was comprised of cells treated with H 2 O 2 and Bleomycin (Bleo). Negative control was comprised of cells treated with CLV. (J) γH2AX immunolocalized to nuclei. For positive control, cells were treated with Bleomycin (Bleo). Intranuclear γH2AX (green) appears as focal spots (red arrows) or as a diffuse pan-nuclear pattern (yellow arrows) in nuclei marked by DAPI (blue) staining. Nuclei are further highlighted by double hashed lines. The boxes in the left pane (scale bars 10 μM) are magnified in the right insets (scale bars 2.5 μm). (K) NFKB1 gene expression. (L) Level of intra-nuclear phosphorylated p65 (p65P). (M) Cellular levels of ROS, IL6, and IL8 were secreted into culture media. (N) Expression levels of senescence markers ( CDKN2A , CDKN1A , CDKN1B , LTA4H , TIMP1 , and MMP1 ). (O) Expression level of LDH toxicity, SAβ-Gal activity, and extracellular level of lactic acid (LA). (P) LDHA1 gene expression. (Q) Rate of proliferation assessed by the quantitation of BrdU incorporated into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. Points are shown as empty and mean points as filled circles. The distribution of data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Positive Control, Incubation, Gene Expression, Activity Assay, Expressing, Negative Control, Staining, Quantitation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET dependent resilience to damage and protection against damage-induced senescence Proliferating aHDFs (0.3 × 10 6 /mL) and PBMCs (70 years; PBMC 70Yr, 1 × 10 6 /mL) were seeded in culture plates and pre-treated in triplicate without (untreated: UT) or with H 2 O 2 (100 μM) for 24 h. After 24 h, cells were washed and treated without or with CLV (20 μM), or CRISPR TET1 ( TET1 CR ;1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression levels of TETs in PBMCs. (B) AKG bioavailability in PBMCs. (C) Expression levels of energy/stress and nutritional sensors in PBMCs. (D) Heatmap of senescence marker gene expression in PBMCs. (E) Level of extracellular lactic acid (LA) and SAβ-Gal activity in PBMCs. (F) Quantification of BrdU and ROS (qROS) in PBMCs. Bar and line graphs show the means ± SD. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 . Points are shown as empty and mean points as filled circles.

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: CRISPR, Incubation, Expressing, Marker, Gene Expression, Activity Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficient senescence state is reversible Young replicatively proliferating aHDFs (week 3, 0.3 × 10 6 /mL)) were seeded in culture plates and pre-treated in triplicates without (UT) and with C35 (5 μM), RLS (20 μM), or TET1 siRNA ( TET1 i ; 300 nM). Aliquot of cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. The remaining cells were re-seeded in equal numbers and cultured without treatment for an additional 7 days (withdrawal). Cultures were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media. (A) Quantification of TET activity, and 5 mC and 5fc levels. (B) Expression level of energy/stress and nutritional sensor genes. (C) Expression level of NFKB1 , RELA , IKBA , COL1A1, and ELN . (D) γH2AX immunolocalized (cyan) in nuclei. Nuclei stained for DAPI (deep blue) are marked by double hashed lines. (E) Extracellular level of lactic acid (LA), SAβ-Gal activity, LDH toxicity, and UPS. (F) Intracellular level of ROS and 8OHdG. (G) Heatmap of senescence markers and RRM2 gene expression. (H) Rate of proliferation assessed by BrdU incorporation into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Incubation, Cell Culture, Activity Assay, Expressing, Staining, Gene Expression, BrdU Incorporation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: Activation of the AKG-TET axis reverses replicative and age induced senescence Replicatively induced senescence. Replicatively senescent aHDFs (0.3 × 10 6 cells/mL) were seeded in triplicate in each plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids (TET1 CR , 1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression level of TET s. (B) AKG bioavailability and TET activity (TET act ). (C) Expression level of energy/stress and nutritional sensor gene expression. (D) Intracellular level of ROS. (E) Heatmap of senescence marker and RRM2 gene expression. (F) Level of lactic acid (LA) in culture media. (G) Cellular SAβ-Gal activity. (H) Quantitation of nuclear BrdU. (I) Total cell numbers. Age induced senescence. PBMCs (PBMC 70Yr ) were seeded (1 × 10 6 cells/mL) in triplicate in each well of a 24-well plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids ( TET1 CR : 1 μg/mL). Cells and culture media were removed on day 7 of treatment for analysis. (J) Quantitation of bioavailable AKG and TET activity in cells treated with RLS versus CLV. (K) qPCR quantitation of the expression level of TET s. (L) qPCR quantitation of the expression level of energy/stress and nutritional sensor gene expression. (M) Quantitation of BrdU incorporated into nuclei of cells in S-phase, lactic (LA) acid in culture media, cellular SAβ-Gal activity, and ROS. (N) Heatmap of senescence markers and RRM2 in PBMCs. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 , ∗∗∗∗p ≤ 5 × 10 −5 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Activation Assay, CRISPR, Incubation, Expressing, Activity Assay, Gene Expression, Marker, Quantitation Assay

Loss of Tet1 exacerbated airway hyperresponsiveness and lung inflammation in a mouse model of asthma. ( a ) Treatment protocol. ( b ) Airway hyperresponsiveness. ( c ) Total BALF cells. ( d ) Number of each cell type. ( e ) Total IgE. ( f ) Total IgG1. ( g ) HDM-specific IgE (1:5 dilution). ( h ) HDM-specific IgG1 (1:2000 dilution). Tet1 +/+ Saline, n = 8–15; Tet1 +/+ HDM, n = 7–24; Tet1 +/− Saline, n = 9–14; Tet1 +/− HDM, n = 9–24; Tet1 −/− Saline, n = 6–7; Tet1 −/− HDM, n = 9–16. mean ± SEM for each group is shown. Unpaired student t-test was applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Loss of Tet1 exacerbated airway hyperresponsiveness and lung inflammation in a mouse model of asthma. ( a ) Treatment protocol. ( b ) Airway hyperresponsiveness. ( c ) Total BALF cells. ( d ) Number of each cell type. ( e ) Total IgE. ( f ) Total IgG1. ( g ) HDM-specific IgE (1:5 dilution). ( h ) HDM-specific IgG1 (1:2000 dilution). Tet1 +/+ Saline, n = 8–15; Tet1 +/+ HDM, n = 7–24; Tet1 +/− Saline, n = 9–14; Tet1 +/− HDM, n = 9–24; Tet1 −/− Saline, n = 6–7; Tet1 −/− HDM, n = 9–16. mean ± SEM for each group is shown. Unpaired student t-test was applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: For the CRISPR plasmid Activation of TET1, control plasmid and TET1 CRISPR activation plasmid (Santa Cruz Biotechnology, Dallas, TX) was transfected into HBECs using Lipofectamine® 2000 (Thermo Fisher Scientific, Florence, KY) and Opti-MEM medium according to manufacturer’s protocol.

Techniques: Saline

Tet1 deficiency increased the expression of pro-Th2/Th17 and epithelial repair response genes. ( a ) Expression levels of indicated genes were measured by RT-qPCR 48 hrs after the last i.t. HDM challenges. Expression values were normalized to the expression of mRpl13a. Mean ± SEM for each group is shown. Tet1 +/+ Saline, n = 4–10; Tet1 +/+ HDM, n = 11–22; Tet1 +/− Saline, n = 4–14; Tet1 +/− HDM, n = 12–21; Tet1 −/− Saline, n = 4–7; Tet1 −/− HDM, n = 10–19. Student t-tests were applied to analyze normally distributed data for Il4 , Il1b, Tslp, Il17f and Egfr with Bonferroni corrections for multiple testing. For comparisons involving data that are not normally distributed in all other genes, Mann Whitney tests were applied. ( b ) Protein levels of IL13, IL5 and IL33 measured by ELISA. IL13 and IL5 were measured in BALF, and IL33 was measured in lung homogenate. Tet1 +/+ Saline, n = 10–11; Tet1 +/+ HDM, n = 8–10; Tet1 +/− Saline, n = 10; Tet1 +/− HDM, n = 10; Tet1 −/− Saline, n = 7; Tet1 −/− HDM, n = 10–11. Mean ± SEM for each group is shown. Data are normally distributed and unpaired student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01. ns represents not significant.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Tet1 deficiency increased the expression of pro-Th2/Th17 and epithelial repair response genes. ( a ) Expression levels of indicated genes were measured by RT-qPCR 48 hrs after the last i.t. HDM challenges. Expression values were normalized to the expression of mRpl13a. Mean ± SEM for each group is shown. Tet1 +/+ Saline, n = 4–10; Tet1 +/+ HDM, n = 11–22; Tet1 +/− Saline, n = 4–14; Tet1 +/− HDM, n = 12–21; Tet1 −/− Saline, n = 4–7; Tet1 −/− HDM, n = 10–19. Student t-tests were applied to analyze normally distributed data for Il4 , Il1b, Tslp, Il17f and Egfr with Bonferroni corrections for multiple testing. For comparisons involving data that are not normally distributed in all other genes, Mann Whitney tests were applied. ( b ) Protein levels of IL13, IL5 and IL33 measured by ELISA. IL13 and IL5 were measured in BALF, and IL33 was measured in lung homogenate. Tet1 +/+ Saline, n = 10–11; Tet1 +/+ HDM, n = 8–10; Tet1 +/− Saline, n = 10; Tet1 +/− HDM, n = 10; Tet1 −/− Saline, n = 7; Tet1 −/− HDM, n = 10–11. Mean ± SEM for each group is shown. Data are normally distributed and unpaired student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01. ns represents not significant.

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

Tet1 deficiency increased the expression of Muc5ac in the lungs. ( a ) Periodic acid–Schiff (PAS) staining of lung sections. Representative images (4 slides per animal, 2–3 animals per group) are shown. ( b ) Expression of Muc5ac ( b ) and Muc5b ( c ) were measured by RT-qPCR. Expression values were normalized to Rpl13a. Mean ± SEM for each group is shown. Tet1 +/+ Saline, n = 8–10; Tet1 +/+ HDM, n = 14–16; Tet1 +/− Saline, n = 12–14; Tet1 +/− HDM, n = 16–21; Tet1 −/− Saline, n = 6–7; Tet1 −/− HDM, n = 15–17. Unpaired student t-tests ( Muc5ac ) or Mann Whitney tests ( Muc5b ) were applied. *p < 0.05. ns represents not significant. Bar represents 100 μm.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Tet1 deficiency increased the expression of Muc5ac in the lungs. ( a ) Periodic acid–Schiff (PAS) staining of lung sections. Representative images (4 slides per animal, 2–3 animals per group) are shown. ( b ) Expression of Muc5ac ( b ) and Muc5b ( c ) were measured by RT-qPCR. Expression values were normalized to Rpl13a. Mean ± SEM for each group is shown. Tet1 +/+ Saline, n = 8–10; Tet1 +/+ HDM, n = 14–16; Tet1 +/− Saline, n = 12–14; Tet1 +/− HDM, n = 16–21; Tet1 −/− Saline, n = 6–7; Tet1 −/− HDM, n = 15–17. Unpaired student t-tests ( Muc5ac ) or Mann Whitney tests ( Muc5b ) were applied. *p < 0.05. ns represents not significant. Bar represents 100 μm.

Techniques: Expressing, Staining, Quantitative RT-PCR, Saline, MANN-WHITNEY

Pathway and protein network analysis of genes differentially expressed between HDM-challenged Tet1 +/+ mice and Tet1 −/− mice. ( a ) Top 11 significantly enriched canonical pathways among the genes that were downregulated in HDM-treated Tet1 −/− mice. ( b ) Significantly enriched canonical pathways among the genes that were upregulated in HDM-treated Tet1 −/− mice. ( c ) Minimal network containing only direct protein-protein interactions between seed proteins from the input dataset. 133 genes differentially expressed genes were used as input, and the links between them indicate known protein-protein interactions. Green color marks genes with downregulated expression, red color marks genes with upregulated expression, grey color marks genes that are not included in the input dataset. ( d ) Significantly enriched pathways among genes included in c. ( e ) RT-qPCR validation of selected differentially expressed genes from RNA-seq. Tet1 +/+ Saline, n = 6–7; Tet1 +/+ HDM, n = 6–7; Tet1 −/− Saline, n = 5–6; Tet1 −/− HDM, n = 7. Expression levels were measured by RT-qPCR and were normalized to the expression of mRpl13a. Mean ± SEM for each group is shown. Data are normally distributed and unpaired student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns represents not significant.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Pathway and protein network analysis of genes differentially expressed between HDM-challenged Tet1 +/+ mice and Tet1 −/− mice. ( a ) Top 11 significantly enriched canonical pathways among the genes that were downregulated in HDM-treated Tet1 −/− mice. ( b ) Significantly enriched canonical pathways among the genes that were upregulated in HDM-treated Tet1 −/− mice. ( c ) Minimal network containing only direct protein-protein interactions between seed proteins from the input dataset. 133 genes differentially expressed genes were used as input, and the links between them indicate known protein-protein interactions. Green color marks genes with downregulated expression, red color marks genes with upregulated expression, grey color marks genes that are not included in the input dataset. ( d ) Significantly enriched pathways among genes included in c. ( e ) RT-qPCR validation of selected differentially expressed genes from RNA-seq. Tet1 +/+ Saline, n = 6–7; Tet1 +/+ HDM, n = 6–7; Tet1 −/− Saline, n = 5–6; Tet1 −/− HDM, n = 7. Expression levels were measured by RT-qPCR and were normalized to the expression of mRpl13a. Mean ± SEM for each group is shown. Data are normally distributed and unpaired student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns represents not significant.

Techniques: Protein-Protein interactions, Expressing, Quantitative RT-PCR, Biomarker Discovery, RNA Sequencing, Saline

IPA predicted direction of change in functions by genes differentially expressed comparing HDM-challenged Tet1 −/− mice with Tet1 +/+ mice.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: IPA predicted direction of change in functions by genes differentially expressed comparing HDM-challenged Tet1 −/− mice with Tet1 +/+ mice.

Techniques: Activation Assay, Infection, Virus, Binding Assay, Migration

Knockdown of TET1 in human bronchial epithelial cells regulates the expression of genes in IFN and AhR pathways. Expression levels of TET1 , IRF7 , IFNα1 , IFNα2 , IFNβ , CYP1A1 , and ALDH1A1 in HBECs with or without TET1 knockdown and treatment of HDM were measured at indicated time points. Expression values of six technical replicates were normalized to the expression of GADPH. Mean ± SEM for each group is shown. Data are normally distributed and student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns represents not significant.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Knockdown of TET1 in human bronchial epithelial cells regulates the expression of genes in IFN and AhR pathways. Expression levels of TET1 , IRF7 , IFNα1 , IFNα2 , IFNβ , CYP1A1 , and ALDH1A1 in HBECs with or without TET1 knockdown and treatment of HDM were measured at indicated time points. Expression values of six technical replicates were normalized to the expression of GADPH. Mean ± SEM for each group is shown. Data are normally distributed and student t-tests were applied with Bonferroni corrections for multiple testing. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns represents not significant.

Techniques: Knockdown, Expressing

Possible mechanisms through which Tet1 regulates IFN and AhR signaling pathways. ( a ) Schematic showing two possible pathways that Tet1 utilizes to silence gene expression. ( b ) DNAm at 5 CpG sites across Irf7 gene (mm10) from RRBS data at p < 0.05. Open bar: Tet1 +/+ HDM; Grey bar: Tet1 −/− HDM. ( c ) Measurement of DNAm levels at CG4 and CG5 in b by bisulfite pyrosequencing. N = 10 for both groups and unpaired student t-tests were applied on normally distributed data. *p < 0.05, **p < 0.01. Open bar: Tet1 +/+ HDM; Grey bar: Tet1 −/− HDM. ( d ) Transcription factor binding motif enrichment in promoter regions of Tet1-regulated genes in human bronchial epithelial cells. The top 30 significantly enriched binding motifs are grouped by their corresponding TFs (p < 10 −2 ). The size of each square or circle indicates fold-enrichment over background. ( e ) Overlap significance between promoter regions (2 kb upstream transcription start site) of Tet1-regulated genes in bronchial epithelial cells and ChIP-seq datasets. The top 30 significant results are shown regardless of cell type, along with results from lung related datasets passing a p < 10 −6 significant threshold. Lung-related cell types are shown in multi-color, and non-lung cell types are shown in dark blue. ( f ) Overlap significance between gene promoters and histone marks. The significance of the degree of overlap between promoters and each member of a large library of histone mark ChIP-seq datasets was estimated. Histone marks with at least one significant result (p < 10 −2 ) are shown. The Y-axis indicates the histone mark, in decreasing order of significance. The X-axis indicates the significance (−log P-value) of the overlap of the given dataset. The size of each circle indicates the fold-enrichment relative to background.

Journal: Scientific Reports

Article Title: TET1 contributes to allergic airway inflammation and regulates interferon and aryl hydrocarbon receptor signaling pathways in bronchial epithelial cells

doi: 10.1038/s41598-019-43767-6

Figure Lengend Snippet: Possible mechanisms through which Tet1 regulates IFN and AhR signaling pathways. ( a ) Schematic showing two possible pathways that Tet1 utilizes to silence gene expression. ( b ) DNAm at 5 CpG sites across Irf7 gene (mm10) from RRBS data at p < 0.05. Open bar: Tet1 +/+ HDM; Grey bar: Tet1 −/− HDM. ( c ) Measurement of DNAm levels at CG4 and CG5 in b by bisulfite pyrosequencing. N = 10 for both groups and unpaired student t-tests were applied on normally distributed data. *p < 0.05, **p < 0.01. Open bar: Tet1 +/+ HDM; Grey bar: Tet1 −/− HDM. ( d ) Transcription factor binding motif enrichment in promoter regions of Tet1-regulated genes in human bronchial epithelial cells. The top 30 significantly enriched binding motifs are grouped by their corresponding TFs (p < 10 −2 ). The size of each square or circle indicates fold-enrichment over background. ( e ) Overlap significance between promoter regions (2 kb upstream transcription start site) of Tet1-regulated genes in bronchial epithelial cells and ChIP-seq datasets. The top 30 significant results are shown regardless of cell type, along with results from lung related datasets passing a p < 10 −6 significant threshold. Lung-related cell types are shown in multi-color, and non-lung cell types are shown in dark blue. ( f ) Overlap significance between gene promoters and histone marks. The significance of the degree of overlap between promoters and each member of a large library of histone mark ChIP-seq datasets was estimated. Histone marks with at least one significant result (p < 10 −2 ) are shown. The Y-axis indicates the histone mark, in decreasing order of significance. The X-axis indicates the significance (−log P-value) of the overlap of the given dataset. The size of each circle indicates the fold-enrichment relative to background.

Techniques: Protein-Protein interactions, Gene Expression, Binding Assay, ChIP-sequencing

Review

Structured Review

Images

1) Product Images from "AKG-TET axis is central to senescence plasticity"

Similar Products

Image Search Results