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Cell Signaling Technology Inc cdkn1a p21
Cdkn1a P21, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1860 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cdkn1a p21 - by Bioz Stars, 2026-07

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a -Transcriptome analyses from P5 hyaloid bulk-RNA sequencing data from R.A. Lang group compared to P5 retina pseudo bulk single-cell RNA sequencing data from S. Blackshaw group. Enrichment analysis was performed on selected mouse gene sets by Gene Set Variation Analysis (GSVA) and coefficient comparison was performed using limma with the difference in GSVA enrichment score (contrast fit coefficient) displayed as bar plot. Blue arrows point senescence-related pathways. b- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) of SASP-related genes ( Tgf-β1, Il1β, Il6, Serpine1 ) comparing between hyaloids and retina at P0, P4 and P8. β-Actin was used as reference gene. Data are presented as fold change compared to retina at each developmental stage (n=3). Bar graphs are means ± SEM. Represented P values are **<0.01, ***<0.001 and **** < 0.0001 from two-tailed parametric unpaired t -test. c - Bar charts of qRT-PCR from SASP-related genes ( Tgf-β1, Il1β, Il6, Serpine1, Vegf-a ) and d- cell cycle arrest markers ( Cdkn1a, Tp53 ) in hyaloids at P0, P4 and P8. Lmnb1 is used as a negative control. β-Actin was used as a reference gene. Data are presented as fold change compared to P0 ( n ≥3-6). Bar graphs are represented as means ± SEM. Represented P values are *<0.05, **<0.01, ***<0.001 and ****<0.0001 from ordinary one-way ANOVA test followed by Dunnett’s multiple comparisons for Tgf-β1, Il1β, Il6, Serpine1, Vegf-a and Cdkn1a and Kruskal-Wallis test followed by Dunn’s multiple comparisons for Tp53 . e - Immunoblots of hyaloid cell lysates from P0, P4 and P8 for senescence markers (P21, TP53, PAI-1). β-ACTIN was used as a loading control (n = 3). f - Representative confocal immunofluorescence staining of P21 (green) and g - PAI-1 (green) of flat-mounted hyaloid vessels at P0, P4 and P8. Blood vessels were stained with IB4 (red). Scale bars, 100 µm. Non-significant (ns).

Journal: bioRxiv

Article Title: Developmental senescence orchestrates hyaloid vessel regression in the postnatal eye

doi: 10.64898/2026.05.12.724389

Figure Lengend Snippet: a -Transcriptome analyses from P5 hyaloid bulk-RNA sequencing data from R.A. Lang group compared to P5 retina pseudo bulk single-cell RNA sequencing data from S. Blackshaw group. Enrichment analysis was performed on selected mouse gene sets by Gene Set Variation Analysis (GSVA) and coefficient comparison was performed using limma with the difference in GSVA enrichment score (contrast fit coefficient) displayed as bar plot. Blue arrows point senescence-related pathways. b- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) of SASP-related genes ( Tgf-β1, Il1β, Il6, Serpine1 ) comparing between hyaloids and retina at P0, P4 and P8. β-Actin was used as reference gene. Data are presented as fold change compared to retina at each developmental stage (n=3). Bar graphs are means ± SEM. Represented P values are **<0.01, ***<0.001 and **** < 0.0001 from two-tailed parametric unpaired t -test. c - Bar charts of qRT-PCR from SASP-related genes ( Tgf-β1, Il1β, Il6, Serpine1, Vegf-a ) and d- cell cycle arrest markers ( Cdkn1a, Tp53 ) in hyaloids at P0, P4 and P8. Lmnb1 is used as a negative control. β-Actin was used as a reference gene. Data are presented as fold change compared to P0 ( n ≥3-6). Bar graphs are represented as means ± SEM. Represented P values are *<0.05, **<0.01, ***<0.001 and ****<0.0001 from ordinary one-way ANOVA test followed by Dunnett’s multiple comparisons for Tgf-β1, Il1β, Il6, Serpine1, Vegf-a and Cdkn1a and Kruskal-Wallis test followed by Dunn’s multiple comparisons for Tp53 . e - Immunoblots of hyaloid cell lysates from P0, P4 and P8 for senescence markers (P21, TP53, PAI-1). β-ACTIN was used as a loading control (n = 3). f - Representative confocal immunofluorescence staining of P21 (green) and g - PAI-1 (green) of flat-mounted hyaloid vessels at P0, P4 and P8. Blood vessels were stained with IB4 (red). Scale bars, 100 µm. Non-significant (ns).

Article Snippet: C57BL/6 wild-type and Cdkn1a (p21) ( Cdkn1a tm1Led /J, no. 016565) knock-out ( Cdkn1a -/- ) [ ] mice lines were obtained from The Jackson Laboratory.

Techniques: RNA Sequencing, Single Cell, Comparison, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Two Tailed Test, Negative Control, Western Blot, Control, Immunofluorescence, Staining

a- Uniform manifold approximation and projection (UMAP) of single-cell RNA sequencing from P4 hyaloid vessels representing the 9 hyaloid cell types identified by graph-based clustering of normalized RNA count. b- Heat map displaying log2 FC of VISION calculated enrichment scores across the 9 hyaloid cell populations for selected senescence- and c - SASP-related gene sets, show enrichment in immune, VSMCs and endothelial cell clusters. d - Dot plot illustrating the expression levels of SenMayo-related genes across the 9 hyaloid cell clusters. e- UMAP feature plot representation of Cdkn1a expression across the 9 hyaloid cell populations. f - Relative mRNA levels of Cdkn1a in P8 hyaloid vessels Cdkn1a +/+ versus Cdkn1a -/- measured by qRT-PCR (n = 3). β-Actin was used as reference gene. g- Representative IB4 and h- SA-β-Gal staining of flat-mounted P8 hyaloid vessels Cdkn1a +/+ (control) and Cdkn1a -/- (lacking Cdkn1a). Higher magnifications of vessels are shown. i - Quantification of total vessel length of P8 flat-mounted hyaloids Cdkn1a +/+ control versus Cdkn1a -/- . Each dot represents a flat-mounted hyaloid sample (n = 8-11). j- Quantification of SA-β-Gal staining per 100 µm of vessels (in percentage) of P8 Cdkn1a +/+ versus Cdkn1a -/- flat-mounted hyaloid. k- Representative IB4 staining of P8 retinal flat-mount Cdkn1a +/+ and Cdkn1a -/- and l - quantification of radial outgrowth of retinal vascular plexus (n = 3), showing delayed expansion of the superficial vascular plexus in Cdkn1a -/- mice. Data are presented as fold change normalized to Cdkn1a +/+ control ( f, j, l ) or as Arbitrary Unit (A.U.) ( i ). Data are represented as means ± SEM ( f, j, l ) or as individual values ( i ). Represented P values are * * < 0.01, *** < 0.001 and **** < 0.0001 from two-tailed parametric unpaired t -test. Scale bars, 500 µm ( g, h ) and 100 µm ( k ) [for higher magnification in g, h ].

Journal: bioRxiv

Article Title: Developmental senescence orchestrates hyaloid vessel regression in the postnatal eye

doi: 10.64898/2026.05.12.724389

Figure Lengend Snippet: a- Uniform manifold approximation and projection (UMAP) of single-cell RNA sequencing from P4 hyaloid vessels representing the 9 hyaloid cell types identified by graph-based clustering of normalized RNA count. b- Heat map displaying log2 FC of VISION calculated enrichment scores across the 9 hyaloid cell populations for selected senescence- and c - SASP-related gene sets, show enrichment in immune, VSMCs and endothelial cell clusters. d - Dot plot illustrating the expression levels of SenMayo-related genes across the 9 hyaloid cell clusters. e- UMAP feature plot representation of Cdkn1a expression across the 9 hyaloid cell populations. f - Relative mRNA levels of Cdkn1a in P8 hyaloid vessels Cdkn1a +/+ versus Cdkn1a -/- measured by qRT-PCR (n = 3). β-Actin was used as reference gene. g- Representative IB4 and h- SA-β-Gal staining of flat-mounted P8 hyaloid vessels Cdkn1a +/+ (control) and Cdkn1a -/- (lacking Cdkn1a). Higher magnifications of vessels are shown. i - Quantification of total vessel length of P8 flat-mounted hyaloids Cdkn1a +/+ control versus Cdkn1a -/- . Each dot represents a flat-mounted hyaloid sample (n = 8-11). j- Quantification of SA-β-Gal staining per 100 µm of vessels (in percentage) of P8 Cdkn1a +/+ versus Cdkn1a -/- flat-mounted hyaloid. k- Representative IB4 staining of P8 retinal flat-mount Cdkn1a +/+ and Cdkn1a -/- and l - quantification of radial outgrowth of retinal vascular plexus (n = 3), showing delayed expansion of the superficial vascular plexus in Cdkn1a -/- mice. Data are presented as fold change normalized to Cdkn1a +/+ control ( f, j, l ) or as Arbitrary Unit (A.U.) ( i ). Data are represented as means ± SEM ( f, j, l ) or as individual values ( i ). Represented P values are * * < 0.01, *** < 0.001 and **** < 0.0001 from two-tailed parametric unpaired t -test. Scale bars, 500 µm ( g, h ) and 100 µm ( k ) [for higher magnification in g, h ].

Article Snippet: C57BL/6 wild-type and Cdkn1a (p21) ( Cdkn1a tm1Led /J, no. 016565) knock-out ( Cdkn1a -/- ) [ ] mice lines were obtained from The Jackson Laboratory.

Techniques: Single Cell, RNA Sequencing, Expressing, Quantitative RT-PCR, Staining, Control, Two Tailed Test

(A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Staining, Western Blot, Expressing, Imaging, Luciferase, Activity Assay, Quantitative RT-PCR

(A, B) Primary human bladder fibroblasts were isolated from surgical excision from bladder cancer patients and subjected to GZMK treatment. (A) HBFs were treated with 100nM GZMK for 5 days, and SA-β-gal staining were performed. (B) HBFs were treated with 100nM GZMK in medium without FBS for one day, IL-6 and TNFα in supernatant were measured by ELISA. (C, D) MEFs were isolated and subjected to GZMK treatment as same with . (E-H) BMDMs were subjected to Gzmk or indicated inhibitors treatment. (E) BMDMs were stimulated with Gzmk for 30 minutes and phosphorylated ERK and p38 were determined by flow cytometry. (F) BMDMs were cultured with Gzmk or combined with indicated inhibitors for three days, SA-β-gal staining were performed. BMDMs were cultured with Gzmk or combined with indicated inhibitors for one day, expression of P16 (G) and P21 (H) were analyzed by flow cytometry, IL-6 and TNFα (I) in supernatant were measured by ELISA.

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A, B) Primary human bladder fibroblasts were isolated from surgical excision from bladder cancer patients and subjected to GZMK treatment. (A) HBFs were treated with 100nM GZMK for 5 days, and SA-β-gal staining were performed. (B) HBFs were treated with 100nM GZMK in medium without FBS for one day, IL-6 and TNFα in supernatant were measured by ELISA. (C, D) MEFs were isolated and subjected to GZMK treatment as same with . (E-H) BMDMs were subjected to Gzmk or indicated inhibitors treatment. (E) BMDMs were stimulated with Gzmk for 30 minutes and phosphorylated ERK and p38 were determined by flow cytometry. (F) BMDMs were cultured with Gzmk or combined with indicated inhibitors for three days, SA-β-gal staining were performed. BMDMs were cultured with Gzmk or combined with indicated inhibitors for one day, expression of P16 (G) and P21 (H) were analyzed by flow cytometry, IL-6 and TNFα (I) in supernatant were measured by ELISA.

Techniques: Isolation, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Expressing

Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Techniques: Clinical Proteomics, Staining, Western Blot

(A-F) Representative immunofluorescence images and corresponding quantitative analysis of senescence and SASP markers (A, B) p21, (C, D) IL-6 and (E, F) TGF-β, n = 4 animals/group, 3 discs/mouse. Scale bar = 50 μm.

Journal: bioRxiv

Article Title: SIRT6 Activation Improves Intervertebral Disc Health in the Aging Spine

doi: 10.64898/2026.03.03.709336

Figure Lengend Snippet: (A-F) Representative immunofluorescence images and corresponding quantitative analysis of senescence and SASP markers (A, B) p21, (C, D) IL-6 and (E, F) TGF-β, n = 4 animals/group, 3 discs/mouse. Scale bar = 50 μm.

Article Snippet: Deparaffinized sections following antigen retrieval or frozen sections were blocked in 5% normal serum in PBS-T, and incubated with antibodies against H3K9Ac (1:50, Sigma, 06-942), Collagen I (1:100, Abcam ab34710), Collagen X (1:500, Abcam ab58632), Aggrecan (1:300, Abcam ab11570); GLUT1 (1:200, Abcam, ab40084), F-CHP (1:100, 3-Helix), p21 (1:200, Novus NB100-1941), IL-6 (1:50, Novus NB600-1131), TGF-ý (1:200, Abcam; ab92486) (n= 4-6 mice/group, 2-3 discs/mouse were analyzed).

Techniques: Immunofluorescence

Transcriptomics identifies HIF1A as a ferroptosis-related hub gene and validates its nuclear translocation in skeletal muscle I/R injury. (A) Volcano plot of differentially expressed genes. (B) Venn diagram of differentially expressed genes and ferroptosis-related genes. (C) GSEA enrichment analysis of ferroptosis-related gene sets in skeletal muscle I/R injury. (D) Bubble plot of KEGG enrichment analysis for 165 ferroptosis-related differentially expressed genes (Fer-DEGs). (E) PPI network of 32 genes in Cluster 1, comprising 32 nodes and 371 edges. (F) PPI network of 6 hub genes ( Hif1a , Cdkn1a , Timp1 , Tlr4 , Cybb , Hmox1 ) within the HIF-1 signaling pathway among the 32 key genes in Cluster 1. (G) Representative immunofluorescence images of HIF1A (red) in C2C12 cells across different experimental groups. Nuclei were counterstained with DAPI (blue). Scale bar represents 100 μm. (H) Quantification of HIF1A immunofluorescence. (I) Cytoplasmic and nuclear expression of HIF1A in C2C12 cells across different experimental groups. (J) Representative immunofluorescence images of HIF1A (red) in skeletal muscle tissue across different experimental groups. Nuclei were counterstained with DAPI (blue). Scale bar represents 100 μm. (K) Quantification of HIF1A immunofluorescence. (L) Cytoplasmic and nuclear expression of HIF1A in skeletal muscle tissue across different experimental groups. Data are expressed as mean ± SD. For in vivo experiments, each group comprised 8-10 animals, with n = 3 randomly selected animals per assay. For in vitro experiments, n = 3 independent experiments. ∗p < 0.05; ∗∗∗∗p < 0.0001.

Journal: Journal of Orthopaedic Translation

Article Title: HIF1A transcriptionally activates CDKN1A to drive ferroptosis in skeletal muscle ischaemia-reperfusion injury

doi: 10.1016/j.jot.2026.101055

Figure Lengend Snippet: Transcriptomics identifies HIF1A as a ferroptosis-related hub gene and validates its nuclear translocation in skeletal muscle I/R injury. (A) Volcano plot of differentially expressed genes. (B) Venn diagram of differentially expressed genes and ferroptosis-related genes. (C) GSEA enrichment analysis of ferroptosis-related gene sets in skeletal muscle I/R injury. (D) Bubble plot of KEGG enrichment analysis for 165 ferroptosis-related differentially expressed genes (Fer-DEGs). (E) PPI network of 32 genes in Cluster 1, comprising 32 nodes and 371 edges. (F) PPI network of 6 hub genes ( Hif1a , Cdkn1a , Timp1 , Tlr4 , Cybb , Hmox1 ) within the HIF-1 signaling pathway among the 32 key genes in Cluster 1. (G) Representative immunofluorescence images of HIF1A (red) in C2C12 cells across different experimental groups. Nuclei were counterstained with DAPI (blue). Scale bar represents 100 μm. (H) Quantification of HIF1A immunofluorescence. (I) Cytoplasmic and nuclear expression of HIF1A in C2C12 cells across different experimental groups. (J) Representative immunofluorescence images of HIF1A (red) in skeletal muscle tissue across different experimental groups. Nuclei were counterstained with DAPI (blue). Scale bar represents 100 μm. (K) Quantification of HIF1A immunofluorescence. (L) Cytoplasmic and nuclear expression of HIF1A in skeletal muscle tissue across different experimental groups. Data are expressed as mean ± SD. For in vivo experiments, each group comprised 8-10 animals, with n = 3 randomly selected animals per assay. For in vitro experiments, n = 3 independent experiments. ∗p < 0.05; ∗∗∗∗p < 0.0001.

Article Snippet: IHC staining with antibodies against HIF1A (20960-1-AP, Proteintech), CDKN1A (10355-1-AP, Proteintech) was performed to measure the protein expression within human skeletal muscle tissues.

Techniques: Transcriptomics, Translocation Assay, Immunofluorescence, Expressing, In Vivo, In Vitro

HIF1A activates Cdkn1a transcription via promoter binding under H/R conditions and correlates with CDKN1A in human skeletal muscle I/R injury. (A) PPI network of 5 hub genes ( Hif1a , Cdkn1a , Timp1 , Tlr4 , Cybb ). (B) Venn diagram illustrating the overlap between 5377 promoter peak-associated genes identified by CUT&Tag sequencing and 5 ferroptosis hub genes. (C) The binding peak of HIF1A within the promoter region of Cdkn1a was visualized by IGV in skeletal muscle I/R injury. (D-E) Analysis of Cdkn1a mRNA and protein levels following Hif1a overexpression. (F-G) Analysis of Cdkn1a mRNA and protein levels following Hif1a knockdown. (H) The ChIP assay results for two specific sites were displayed on agarose gels, with the samples categorized into Input, IgG, and HIF1A groups in C2C12 cells subjected to H/R conditions. Additionally, densitometric quantification of the ChIP assay was performed. (I) Dual-luciferase reporter assay in 293T cells co-transfected with Hif1a overexpression plasmid and Cdkn1a-WT or Cdkn1a-MUT promoter constructs. Firefly/Renilla luciferase activity ratio was normalized to the empty vector control. Data are expressed as mean ± SD. n = 3 independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, non-significant. (J) Representative immunohistochemical images of HIF1A and CDKN1A protein expression in normal and I/R skeletal muscle. Scale bar: 100 μm. (K) Quantification of HIF1A and CDKN1A immunostaining intensity; ∗∗p < 0.01; ∗∗∗p < 0.001. (L) Spearman correlation analysis between HIF1A and CDKN1A protein expression levels across all muscle samples (r = 0.543, p = 0.006; n = 12 patients).

Journal: Journal of Orthopaedic Translation

Article Title: HIF1A transcriptionally activates CDKN1A to drive ferroptosis in skeletal muscle ischaemia-reperfusion injury

doi: 10.1016/j.jot.2026.101055

Figure Lengend Snippet: HIF1A activates Cdkn1a transcription via promoter binding under H/R conditions and correlates with CDKN1A in human skeletal muscle I/R injury. (A) PPI network of 5 hub genes ( Hif1a , Cdkn1a , Timp1 , Tlr4 , Cybb ). (B) Venn diagram illustrating the overlap between 5377 promoter peak-associated genes identified by CUT&Tag sequencing and 5 ferroptosis hub genes. (C) The binding peak of HIF1A within the promoter region of Cdkn1a was visualized by IGV in skeletal muscle I/R injury. (D-E) Analysis of Cdkn1a mRNA and protein levels following Hif1a overexpression. (F-G) Analysis of Cdkn1a mRNA and protein levels following Hif1a knockdown. (H) The ChIP assay results for two specific sites were displayed on agarose gels, with the samples categorized into Input, IgG, and HIF1A groups in C2C12 cells subjected to H/R conditions. Additionally, densitometric quantification of the ChIP assay was performed. (I) Dual-luciferase reporter assay in 293T cells co-transfected with Hif1a overexpression plasmid and Cdkn1a-WT or Cdkn1a-MUT promoter constructs. Firefly/Renilla luciferase activity ratio was normalized to the empty vector control. Data are expressed as mean ± SD. n = 3 independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, non-significant. (J) Representative immunohistochemical images of HIF1A and CDKN1A protein expression in normal and I/R skeletal muscle. Scale bar: 100 μm. (K) Quantification of HIF1A and CDKN1A immunostaining intensity; ∗∗p < 0.01; ∗∗∗p < 0.001. (L) Spearman correlation analysis between HIF1A and CDKN1A protein expression levels across all muscle samples (r = 0.543, p = 0.006; n = 12 patients).

Techniques: Binding Assay, Sequencing, Over Expression, Knockdown, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Construct, Activity Assay, Control, Immunohistochemical staining, Expressing, Immunostaining

Inhibiting HIF1A mitigates lipid peroxidation and ferroptosis triggered by H/R in C2C12 cells by regulating the transcription of Cdkn1a . (A) Cell inhibition rate detected by CCK8 assay. (B) GSH level. (C) MDA level. (D) ROS level. (E) Iron level. (F-L) The mRNA and protein levels of GPX4, ACSL4, PTGS2. Data are expressed as mean ± SD. n = 3 independent experiments. ∗∗∗∗p < 0.0001.

Journal: Journal of Orthopaedic Translation

Article Title: HIF1A transcriptionally activates CDKN1A to drive ferroptosis in skeletal muscle ischaemia-reperfusion injury

doi: 10.1016/j.jot.2026.101055

Figure Lengend Snippet: Inhibiting HIF1A mitigates lipid peroxidation and ferroptosis triggered by H/R in C2C12 cells by regulating the transcription of Cdkn1a . (A) Cell inhibition rate detected by CCK8 assay. (B) GSH level. (C) MDA level. (D) ROS level. (E) Iron level. (F-L) The mRNA and protein levels of GPX4, ACSL4, PTGS2. Data are expressed as mean ± SD. n = 3 independent experiments. ∗∗∗∗p < 0.0001.

Techniques: Inhibition, CCK-8 Assay

CDKN1A suppression rescues ferroptosis in skeletal muscle I/R injury across cellular and animal models. (A) Effect of Cdkn1a knockdown on the inhibition rate of H/R treated C2C12 cells detected by CCK8 assay. (B) GSH level. (C) MDA level. (D) ROS level. (E) Iron level. (F-I) The protein levels of GPX4, ACSL4, PTGS2. (J) Representative HE-stained photomicrographs of gastrocnemius muscle sections from Sham, I/R, Sham + UC2288, and I/R + UC2288 experimental groups. Neutrophil infiltration was observed following reperfusion, and this infiltration was alleviated with UC2288 treatment. (K) Histological injury score. (L) Skeletal muscle wet/dry weight ratio. (M) GSH level in skeletal muscle tissues. (N) MDA level in skeletal muscle tissues. (O) Infarct ratio. (P) ROS level in skeletal muscle tissues. (Q) Iron level in skeletal muscle tissues. (R) Photographs of TTC-stained skeletal muscle sections of different groups. (S-V) The protein levels of GPX4, ACSL4, PTGS2. Data are expressed as mean ± SD. For in vivo experiments, each group comprised 8-10 animals, with n = 3 randomly selected animals per assay. For in vitro experiments, n = 3 independent experiments. ∗∗∗∗p < 0.0001.

Journal: Journal of Orthopaedic Translation

Article Title: HIF1A transcriptionally activates CDKN1A to drive ferroptosis in skeletal muscle ischaemia-reperfusion injury

doi: 10.1016/j.jot.2026.101055

Figure Lengend Snippet: CDKN1A suppression rescues ferroptosis in skeletal muscle I/R injury across cellular and animal models. (A) Effect of Cdkn1a knockdown on the inhibition rate of H/R treated C2C12 cells detected by CCK8 assay. (B) GSH level. (C) MDA level. (D) ROS level. (E) Iron level. (F-I) The protein levels of GPX4, ACSL4, PTGS2. (J) Representative HE-stained photomicrographs of gastrocnemius muscle sections from Sham, I/R, Sham + UC2288, and I/R + UC2288 experimental groups. Neutrophil infiltration was observed following reperfusion, and this infiltration was alleviated with UC2288 treatment. (K) Histological injury score. (L) Skeletal muscle wet/dry weight ratio. (M) GSH level in skeletal muscle tissues. (N) MDA level in skeletal muscle tissues. (O) Infarct ratio. (P) ROS level in skeletal muscle tissues. (Q) Iron level in skeletal muscle tissues. (R) Photographs of TTC-stained skeletal muscle sections of different groups. (S-V) The protein levels of GPX4, ACSL4, PTGS2. Data are expressed as mean ± SD. For in vivo experiments, each group comprised 8-10 animals, with n = 3 randomly selected animals per assay. For in vitro experiments, n = 3 independent experiments. ∗∗∗∗p < 0.0001.

Techniques: Knockdown, Inhibition, CCK-8 Assay, Staining, In Vivo, In Vitro

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