nfatc1 abclonal antibody  (ABclonal Biotechnology)

Journal: Bone Research

Article Title: Human peripheral osteoclast-precursor-development patterns reveal the significance of RPS17-dependent ribosome synthesis to Ankylosing Spondylitis lesions

doi: 10.1038/s41413-025-00474-5

Figure Lengend Snippet: Single-monocyte transcriptional profiling and identification of monocytic OCPs. a Schematic diagram showing the overall study design. The scRNA-seq was based on PBMCs across four conditions and the output data were applied for sc-transcriptome analysis and primitive clustering. Five out of 23 clusters that fit the genetic characteristics of monocytes were reclustered and subjected to sc-transcriptome analysis again. b UMAP projection based on monocytic types, and each dot is colored according to cell subtype. c Gating strategy for monocyte enrichment and clustering. Monocytes (CD14 + CD3 − CD19 − NKp46 − cells) were enriched from PBMCs, and four specific monocyte populations were sorted from the CD14 + gate based on CD14 and CD16 expression using FACS. d Osteoclastogenic capacities of various monocytic clusters (CD14 bright CD16 − , CD14 dim CD16 + , CD14 bright CD16 + , CD14 dim CD16 − and CD14 + CD16 − cells) and total monocytes. Scale bar: 100 µm. e Western-blotting analysis of osteoclastic markers (TRAP, CTSK, MMP9 and NFATC1) among five monocytic clusters and total monocytes. The relative levels of each protein are expressed as the ratio of the target protein to GAPDH. f Merged cell-trajectories of mono subsets 0/1/2/5/7/9 (Monocle-2). Different dots correspond to different subsets. g Pseudotimeline of cell-trajectories. The deeper dot color indicates a smaller pseudotime, i.e., an earlier development stage. The changes from a to b/from b to c/from c to d indicate a significant decrease with P < 0.05, and double-letter indicates no statistical difference between the given group and the compared groups marked with a single letter

Article Snippet: After blockage with 5% milk, PVDFs were combined with antibodies targeting GAPDH (1:20 000, 10494-1-AP, Proteintech; IL, USA), TRAP (1:1 000, 11594-1-AP, Proteintech), CTSK (1:400, A1782, ABclonal; Wuhan, China), MMP9 (1:1 000, A0289, ABclonal), NFATC1 (1:1 000, A19597, ABclonal), CD14 (1:1 000, 17000-1-AP, Proteintech), S100A8 (1:1 000, 15792-1-AP, Proteintech), S100A9 (1:2 000, 26992-1-AP, Proteintech), S100A12 (1:1 000, A5328, ABclonal), RPLP1 (1:1 500, A6725, ABclonal), RPL13 (1:1 000, A9404, ABclonal), RPS12 (1:1 500, A5890, ABclonal), RPS28 (1:1 500, A17937, ABclonal), RPS17 (1:1 500, 16267-1-AP, Proteintech), MT-ND6 (1:2 000, A17991, ABclonal), NAMPT (1:2 000, A0256, ABclonal), CSTA (1:2 000, bsm-62286R, Bioss) and CFD (1:2 000, bs-13130R, Bioss) at 4 °C overnight.

Techniques: Expressing, Western Blot

Journal: Bone Research

Article Title: Human peripheral osteoclast-precursor-development patterns reveal the significance of RPS17-dependent ribosome synthesis to Ankylosing Spondylitis lesions

doi: 10.1038/s41413-025-00474-5

Figure Lengend Snippet: AS-associated monocytic evolution towards OCP-lineage fate. a Merged cell-trajectories of the above six subsets showing 5 cell-states and 2 cell-branches. State-1 is the starting point for differentiation. b Bubble plots showing the expression distribution of top genes belonging to various cell-states across five states. Circle size indicates percentage of cells expressing corresponding gene (PCT) and color depth indicates average expression levels. c GSVA scores in the osteoclastogenic functions between five states. Rows were normalized, and the transition of blue-white-red indicates an increase in the scores. d Heatmap showing relative intergroup comparisons of ACP5 , NFATC1 and OSCAR expression between five states. Rows were normalized, and the transition of blue-white-red indicates an increase in the gene expression. e PCT of ACP5 , NFATC1 and OSCAR across five states. f Immunofluorescence analysis of co-expression of MT-ND6, NAMPT and CSTA with OSCAR in CD14 + CD16 − monocytes. M + O + Cells/M + Cells represents the ratio of MT-ND6 and OSCAR-double-positive cells to MT-ND6-positive cells; N + O + Cells/N + Cells represents the ratio of NAMPT and OSCAR-double-positive cells to NAMPT-positive cells; C + O + Cells/C + Cells represents the ratio of CSTA and OSCAR-double-positive cells to CSTA-positive cells. g – i Flow cytometry of CD14 + CD16 − monocytes identifying the approximate trend of MT-ND6 + , NAMPT + and CSTA + cells at the 2nd, 4th, 8th and 16th hours of osteoclastic induction. j Proportions of five states from four groups. k Osteoclast differentiation capacities for CD14 + CD16 − monocytes of each three subjects from four groups. Scale bar: 100 µm. Data come from the mean values of osteoclasts in each three wells. l The expression distribution of MT-ND6, NAMPT, CSTA and CFD across four groups. Circle size indicates PCT and color depth indicates average levels. m Western-blotting analysis of MT-ND6, NAMPT, CSTA and CFD for CD14 + CD16 − monocytes from four groups of subjects ( n = 6/group; each 2 samples presented in one group). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1 and ns (not significant) by one-way ANOVA

Article Snippet: After blockage with 5% milk, PVDFs were combined with antibodies targeting GAPDH (1:20 000, 10494-1-AP, Proteintech; IL, USA), TRAP (1:1 000, 11594-1-AP, Proteintech), CTSK (1:400, A1782, ABclonal; Wuhan, China), MMP9 (1:1 000, A0289, ABclonal), NFATC1 (1:1 000, A19597, ABclonal), CD14 (1:1 000, 17000-1-AP, Proteintech), S100A8 (1:1 000, 15792-1-AP, Proteintech), S100A9 (1:2 000, 26992-1-AP, Proteintech), S100A12 (1:1 000, A5328, ABclonal), RPLP1 (1:1 500, A6725, ABclonal), RPL13 (1:1 000, A9404, ABclonal), RPS12 (1:1 500, A5890, ABclonal), RPS28 (1:1 500, A17937, ABclonal), RPS17 (1:1 500, 16267-1-AP, Proteintech), MT-ND6 (1:2 000, A17991, ABclonal), NAMPT (1:2 000, A0256, ABclonal), CSTA (1:2 000, bsm-62286R, Bioss) and CFD (1:2 000, bs-13130R, Bioss) at 4 °C overnight.

Techniques: Expressing, Gene Expression, Immunofluorescence, Flow Cytometry, Western Blot

Journal: Lasers in Medical Science

Article Title: Effects of low level laser on periodontal tissue remodeling in hPDLCs under tensile stress

doi: 10.1007/s10103-023-03885-0

Figure Lengend Snippet: Gene sequence

Article Snippet: The primary antibodies were listed in the following manner: GAPDH (ABclonal,1:5000,USA), NFATC1 (ABclonal,1:1000,USA), TIMP-1 (ABclonal,1:1000,USA),TIMP-2 (ABclonal,1:1000,USA), MMP-1 (ABclonal,1:1000,USA),MMP-2 (ABclonal,1:1000,USA), Runx2 (ABclonal,1:1000,USA),MMP-9 (ABclonal,1:1000,USA), COL-I (ABclonal,1:1000,USA), COL-III (ABclonal,1:1000,USA), c-Fos (CST, 1:1000, US), OCN (CST, 1:1000), CTSK (Abcam,1:1000).

Techniques: Sequencing

Journal: Lasers in Medical Science

Article Title: Effects of low level laser on periodontal tissue remodeling in hPDLCs under tensile stress

doi: 10.1007/s10103-023-03885-0

Figure Lengend Snippet: Osteogenic differentiated factors and Osteoclast differentiated factors detection ( a ) Quantitative real-time PCR analysis of Runx2 and OCN mRNA expression of hPDLCs. n = 3 . ( b ) Western blot analysis of Runx2 and OCN protein levels of hPDLCs. n = 3 . ( c ) Quantitative real-time PCR analysis of NFATC1, c-Fos and CTSK mRNA expression of hPDLCs. n = 3 . ( d ) Western blot analysis of NFATC1, C-FOS and CTSK protein levels of hPDLCs. n = 3 . Data represent the mean ± SD of three independent experiments. *P < 0.05 vs. the control group, #P < 0.05 vs. the tensile stress group and &P < 0.05 vs. the laser group by one-way ANOVA

Article Snippet: The primary antibodies were listed in the following manner: GAPDH (ABclonal,1:5000,USA), NFATC1 (ABclonal,1:1000,USA), TIMP-1 (ABclonal,1:1000,USA),TIMP-2 (ABclonal,1:1000,USA), MMP-1 (ABclonal,1:1000,USA),MMP-2 (ABclonal,1:1000,USA), Runx2 (ABclonal,1:1000,USA),MMP-9 (ABclonal,1:1000,USA), COL-I (ABclonal,1:1000,USA), COL-III (ABclonal,1:1000,USA), c-Fos (CST, 1:1000, US), OCN (CST, 1:1000), CTSK (Abcam,1:1000).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control