primary antibodies against p21  (Santa Cruz Biotechnology)

Journal: Scientific Reports

Article Title: Vibrational spectroscopy unveils distinct cell cycle features of cancer stem cells in melanoma

doi: 10.1038/s41598-025-14018-8

Figure Lengend Snippet: Immunohistochemical staining of p16 and p21 expression in CHL-1 melanoma cells, CD133 + melanoma stem-like cells, and CD133- non-stem melanoma cells at different time points (11 h, 24 h, 48 h, and 72 h). Representative images show the time-dependent expression of p16 and p21 in each cell type, with higher magnification insets. Scale bars: 100 μm. Bar graphs on the right represent the quantification of p16- and p21-positive cells (mean ± SD) for each group at the indicated time points.

Article Snippet: Primary antibodies against p21 (Bioss, BB07142233) and p16 (Bioss, BB10114929) were diluted at 1:200 and applied to the cells.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Bioactive Materials

Article Title: Liver-bone organoids reveal senescence-driven interorgan crosstalk

doi: 10.1016/j.bioactmat.2025.08.039

Figure Lengend Snippet: Senescent bone organoids construction by replication. IHC staining of SA-β-gal in bone tissues (A) and primary BMSCs (B) of 4-month-old and 14-month-old mice. Scale bars, 200 μm and 100 μm. (C) Diagram of senescent bone organoids construction via 3D bioprinting. (D) Proliferation of P3 and P23 BMSCs. (E) IF staining of SA-β-gal in P3 and P23 BMSCs using a fluorescent probe (red). Scale bars, 100 μm. (F) IF staining of p21 (green), SA-β-gal (red), γ-H2A.X (purple) and DAPI (blue) in bone organoids constructed by P3 and P23 BMSCs. Scale bars, 200 μm. (G) Mineralization of bone organoids assessed by micro-CT scanning. (H–J) Changes and quantitative analysis of BMD (H), BV/TV (I) and Tb.Th (J). Data are presented as the mean ± SD. Data are considered statistically significant at ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against p21 (1:200, CST, 64016), γ-H2A.X (1:200, CST, 80312) were applied overnight (4 °C).

Techniques: Immunohistochemistry, Staining, Construct, Micro-CT

Journal: Bioactive Materials

Article Title: Liver-bone organoids reveal senescence-driven interorgan crosstalk

doi: 10.1016/j.bioactmat.2025.08.039

Figure Lengend Snippet: Senescent bone organoids construction by inducer. (A) IHC staining of SA-β-gal in primary BMSCs treated with 0.5 μM DOX for 48 h. Scale bars, 50 μm. (B) Diagram of DOX-induced senescent bone organoids construction. (C–F) Represented images of 3D mineralization of bone organoids treated with 0.5 μM DOX for 3 and 6days (C) and quantification of BMD (D), BV/TV (E) and Tb.Th (F). (G–H) IF staining of p21 (green), SA-β-gal (red), γ-H2A.X (purple) and DAPI (blue) in bone organoids treated with 0.5 μM DOX (G), D-gal and H 2 O 2 (H). Scale bars, 200 μm. (I–L) Represented images of 3D mineralization of bone organoids treated with D-gal and H 2 O 2 for 6days (I) and quantification of BMD (J), BV/TV (K) and Tb.Th (L). Data are presented as the mean ± SD. Data are considered statistically significant at ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against p21 (1:200, CST, 64016), γ-H2A.X (1:200, CST, 80312) were applied overnight (4 °C).

Techniques: Immunohistochemistry, Staining

Journal: Bioactive Materials

Article Title: Liver-bone organoids reveal senescence-driven interorgan crosstalk

doi: 10.1016/j.bioactmat.2025.08.039

Figure Lengend Snippet: Construction of senescent liver organoids. (A) IF staining of SA-β-gal in liver tissues of 4-month-old and 14-month-old mice. Scale bars, 200 μm. (B) Diagram of senescent liver organoids construction from aged mice. (C) IF staining of SA-β-gal in liver organoids constructed from the duct of 4-month-old and 14-month-old mice. Scale bars, 100 μm. (D) SA-β-gal detection in AML12 cells treated with 0.5 μM DOX for 24 h. Scale bars, 50 μm. (E) The protein level of p53, p21 and p16. (F) Relative mRNA levels of p21. (G) Diagram of DOX-induced senescent liver organoids construction. (H) IF staining of p21 (green), SA-β-gal (red), γ-H2A.X (purple) and DAPI (blue) in liver organoids treated with 0.5 μM DOX for 3 and 6 days. Scale bars, 100 μm. (I) Lipid accumulation of liver organoids. Scale bars, 100 μm. Data are presented as the mean ± SD. Data are considered statistically significant at ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against p21 (1:200, CST, 64016), γ-H2A.X (1:200, CST, 80312) were applied overnight (4 °C).

Techniques: Staining, Construct

Journal: Bioactive Materials

Article Title: Liver-bone organoids reveal senescence-driven interorgan crosstalk

doi: 10.1016/j.bioactmat.2025.08.039

Figure Lengend Snippet: Senescence propagation between liver and bone organoids. (A–B) IF staining of senescence markers in liver organoids (SA-β-gal) (A) and bone organoids (p21, SA-β-gal and γ-H2A.X) (B) treated with 10 % serum (excluded FBS) from 4-month-old and 14-month-old mice. Scale bars, 100 μm and 200 μm. (C–F) Represented images of 3D mineralization (C) and quantification of BMD (D), BV/TV (E) and Tb.Th (F). (G) Diagram of the senescent conditioned medium exchange between liver and bone organoids. (H) IF staining of SA-β-gal in liver organoids treated with conditioned medium from senescent bone organoids. Scale bars, 100 μm. (I) IF staining of p21 (green), SA-β-gal (red), γ-H2A.X (purple) and DAPI (blue) in bone organoids treated with conditioned medium from senescent liver organoids. Scale bars, 200 μm. (J–M) Represented images of 3D mineralization (J) and quantification of BMD (K), BV/TV (L) and Tb.N (M). Data are presented as the mean ± SD. Data are considered statistically significant at ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against p21 (1:200, CST, 64016), γ-H2A.X (1:200, CST, 80312) were applied overnight (4 °C).

Techniques: Staining

Journal: Bioactive Materials

Article Title: Liver-bone organoids reveal senescence-driven interorgan crosstalk

doi: 10.1016/j.bioactmat.2025.08.039

Figure Lengend Snippet: 27-hydroxycholesterol drives senescence propagation to bone organoids. (A) IF staining of senescence markers (p21, SA-β-gal and γ-H2A.X) in bone organoids treated with 2.5 μM 27- OHC for 6 days. Scale bars, 200 μm. (B–E) Represented images of 3D mineralization of bone organoids treated with DOX and 27-OHC (B) and quantitative analysis of BMD (C), BV/TV (D) and Tb.Th (E). (F) H&E, Masson's Trichrome and ARS staining of bone organoids treated with DOX and 27-OHC. Scale bars, 200 μm. 10-week-old male mice were administered the 27-OHC via intraperitoneal injection daily for 20 days after HDT PT3-NRAS and AKT along with transposase ( SB100 ) (n = 5 per group). (G–K) Represented images of 3D scanning (G) and quantification of BMD (H), BV/TV (I), Tb.N (J), and Tb.Sp (K). Data are presented as the mean ± SD. Data are considered statistically significant at ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against p21 (1:200, CST, 64016), γ-H2A.X (1:200, CST, 80312) were applied overnight (4 °C).

Techniques: Staining, Injection

Journal: Aging Cell

Article Title: Mechanistic Insights Into 5′‐ tiRNA ‐His‐ GTG Mediated Activation of the JNK Pathway in Skin Photoaging

doi: 10.1111/acel.70049

Figure Lengend Snippet: Overexpression of 5′‐tiRNA‐His‐GTG in HDF cells induced cellular senescence. (A) qRT‐PCR validation of the overexpression of 5′‐tiRNA‐His‐GTG in HDF cells. (B) Overexpressing 5′‐tiRNA‐His‐GTG in HDF cells and staining with SA‐β‐gal after 24 h. Cells with blue staining represent senescent cells. The treated cells were divided into three groups: unstained, strongly positive, and weakly positive. The percentage of cells in each group was presented in a bar graph. The scale bar, 100 μm. (C) WB analysis of Collagen Type I, p53, and p21 in HDF cells after 5′‐tiRNA‐His‐GTG Mimic transfection. (D) The mRNA levels of IL ‐ 1β , IL ‐ 6 , and IL ‐ 8 in HDF cells after 5′‐tiRNA‐His‐GTG Mimic transfection. (E) EdU assay was used to analyze HDF cells' proliferation ability after overexpressing 5′‐tiRNA‐His‐GTG. The scale bar is 100 μm.

Article Snippet: Primary antibodies against p21 (10355‐1‐AP), p53 (10442‐1‐AP), and Collagen type I polyclonal antibody (14695‐1‐AP) were purchased from Proteintech (China).

Techniques: Over Expression, Quantitative RT-PCR, Biomarker Discovery, Staining, Transfection, EdU Assay

Journal: Aging Cell

Article Title: Mechanistic Insights Into 5′‐ tiRNA ‐His‐ GTG Mediated Activation of the JNK Pathway in Skin Photoaging

doi: 10.1111/acel.70049

Figure Lengend Snippet: Inhibiting 5′‐tiRNA‐His‐GTG rescued UVB‐induced HDF cells photoaging. (A) HDF cells were exposed to UVB after inhibiting 5′‐tiRNA‐His‐GTG, and stained with SA‐β‐gal after 24 h. Cells with blue staining represent senescent cells. The treated cells were divided into three groups: unstained, strongly positive, and weakly positive. The percentage of cells in each group was presented in a bar graph. The scale bar, 100 μm. (B) WB analysis of Collagen Type I, p53, and p21 in the photoaging cell model after 5′‐tiRNA‐His‐GTG Inhibitor transfection. (C) The mRNA levels of IL ‐ 1β , IL ‐ 6 , and IL ‐ 8 in the photoaging cell model after 5′‐tiRNA‐His‐GTG Inhibitor transfection. (D) EdU assay was used to analyze cell proliferation ability in the photoaging cell model after 5′‐tiRNA‐His‐GTG Inhibitor transfection. The scale bar, 100 μm.

Article Snippet: Primary antibodies against p21 (10355‐1‐AP), p53 (10442‐1‐AP), and Collagen type I polyclonal antibody (14695‐1‐AP) were purchased from Proteintech (China).

Techniques: Staining, Transfection, EdU Assay

Journal: Aging Cell

Article Title: Mechanistic Insights Into 5′‐ tiRNA ‐His‐ GTG Mediated Activation of the JNK Pathway in Skin Photoaging

doi: 10.1111/acel.70049

Figure Lengend Snippet: Overexpression of NUP98 rescues 5′‐tiRNA‐His‐GTG‐induced cell senescence in HDF cells. (A) WB analysis of oe‐NUP98 in HDF cells. (B) WB analysis of Collagen Type I, p53 and p21 in HDF cells and oe‐NUP98 HDF cells, with 5′‐tiRNA‐His‐GTG Mimic transfection. (C) The mRNA levels of IL ‐ 1β , IL ‐ 6 and IL ‐ 8 in HDF cells and oe‐NUP98 HDF cells, after 5′‐tiRNA‐His‐GTG Mimic transfection. (D) EdU assay was used to analyze HDF cells and oe‐NUP98 HDF cells proliferation ability after 5′‐tiRNA‐His‐GTG Mimic transfection. The scale bar, 100 μm. (E) HDF cells and oe‐NUP98 HDF cells were transfected with 5′‐tiRNA‐His‐GTG Mimic and stained with SA‐β‐gal after 48 h. Cells with blue staining represent senescent cells. The treated cells were divided into three groups: unstained, strongly positive, and weakly positive. The percentage of cells in each group was presented in a bar graph. The scale bar, 100 μm.

Article Snippet: Primary antibodies against p21 (10355‐1‐AP), p53 (10442‐1‐AP), and Collagen type I polyclonal antibody (14695‐1‐AP) were purchased from Proteintech (China).

Techniques: Over Expression, Transfection, EdU Assay, Staining

Journal: Aging Cell

Article Title: Mechanistic Insights Into 5′‐ tiRNA ‐His‐ GTG Mediated Activation of the JNK Pathway in Skin Photoaging

doi: 10.1111/acel.70049

Figure Lengend Snippet: Overexpression of NUP98 attenuates cellular senescence. (A) oe‐NUP98 HDF cells were stained with SA‐β‐gal after 48 h. Cells with blue staining represent senescent cells. The treated cells were divided into three groups: unstained, strongly positive, and weakly positive. The percentage of cells in each group was presented in a bar graph. The scale bar is 100 μm. (B) The mRNA levels of IL ‐ 1β , IL ‐ 6 and IL ‐ 8 in oe‐NUP98 HDF cells. (C) WB analysis of NUP98, Collagen Type I, p53 and p21 in oe‐NUP98 HDF cells. (D) EdU assay was used to analyze oe‐NUP98 HDF cells' proliferation ability. The scale bar is 100 μm.

Article Snippet: Primary antibodies against p21 (10355‐1‐AP), p53 (10442‐1‐AP), and Collagen type I polyclonal antibody (14695‐1‐AP) were purchased from Proteintech (China).

Techniques: Over Expression, Staining, EdU Assay