primary anti p21  (Proteintech)

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: Materials Today Bio

Article Title: Modeling a mesenchymal cell state by bioprinting for the molecular analysis of dormancy in melanoma

doi: 10.1016/j.mtbio.2025.101674

Figure Lengend Snippet: CIB culture induces a stable mesenchymal phenotype of Mel Im. Assignment of DEGs comparing CIB- and MG-cultured cells with published scRNA-Seq data of malignant melanoma . The cell state annotated marker genes described by Pozniak et al. were used to match the CIB specific gene signature composed of upregulated genes (LFC >1.5) with the validated cell clusters in human melanoma tumor biopsy samples. A) Significantly upregulated genes in CIB cultivated cells showed an enrichment in the neural crest like and mesenchymal defined cell clusters and B) significantly upregulated genes under MG culture conditions illustrate an enrichment in the mitotic cell cluster. C) Expression status of the respective marker genes of the mitotic, neural crest like and mesenchymal cell type clusters in CIB- and MG-cultured melanoma cells based on the scaled normalized RNA-Seq count data. D) CDKN1A RNA-Seq read counts (normalized to library size) in CIB- and MG-cultured Mel Im cells. E) CDKN1A (p21) mRNA expression analysis of Mel Im cultured in CIB or MG by qRT-PCR. F) Immunohistochemical staining for p21 on fixed and paraffin-embedded sections of CIB- or MG-cultured Mel Im. Scale bar = 100 μm. G) Representative images and quantification of Mel Im FUCCI after siPool mediated knockdown of p21 in CIB culture. Scale bar = 100 μm. H) KLF4 RNA-Seq read counts (normalized to library size) in CIB- and MG-cultured Mel Im cells. I) KLF4 mRNA expression analysis of Mel Im cultured in CIB or MG by qRT-PCR. J) Immunohistochemical staining for KLF4 on fixed and paraffin-embedded sections of CIB- or MG-cultured Mel Im. Scale bar = 100 μm. K) Representative images and quantification of Mel Im FUCCI after siPool mediated knockdown of KLF4 in CIB culture. Scale bar = 100 μm ∗p ≤ 0.05 ( D , E , H , I : Student's t-test. G , K : Two-way ANOVA followed by Bonferroni post-test).

Article Snippet: Immunohistochemical staining was performed as described previously, using primary antibodies anti-p21 (1:50, #2947, Cell Signaling, Danvers, MA, USA) or anti-KLF4 (1:500, HPA002926, Sigma Aldrich, St. Louis, MO, USA) [ ].

Techniques: Cell Culture, Marker, Expressing, RNA Sequencing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Knockdown

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