cell signaling technology hsp90  (Cell Signaling Technology Inc)

Journal: Journal of Translational Medicine

Article Title: Sinensetin serves as an AMPK activator to inhibit RANKL-induced osteoclastogenesis via osteoclast cytoskeleton reorganization

doi: 10.1186/s12967-025-06708-8

Figure Lengend Snippet: SIN prevents the cytoskeleton reorganization of preosteoclasts. a Schematic diagram of osteoclast differentiation and maturation. b Images of crystalline violet staining for the transwell migration assay were recorded. c Statistical area ratio of invaded cells. d Images of cells at the same scratch location after 0 and 24 h in the presence and absence of SIN. e The number of cells in the same region were counted and quantified the difference in number between 0 and 24 h. f Representative images of western blot showed the expression of c-Src, Integrin β3 and Cortactin standardized to β-actin. g – i Quantitative analysis of c-Src, Integrin β3 and Cortactin normalized to β-actin (n = 3). j Representative images of western blot showed the expression of p-PAK4, p-PI3K and p-AKT individually standardized to β-actin, PI3K and AKT. (K-M) Quantitative analysis of p-PAK4, p-PI3K and p-AKT proteins expression (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001. All data are expressed as mean ± SD

Article Snippet: Primary antibodies against c-Fos (ab134122), ATP6V0D2 (ab236375), p-AMPK (ab133448) and AMPK (ab32047) were obtained from Abcam (Cambridge, UK), NFATc1 (sc-7294) and CTSK (sc-48353) were from Santa Cruz Biotechnology (Dallas, USA), p-PAK4 (bs-2270R) were obtained from Bioss (Beijing, China), Cortactin ( R23963 ) were obtained from ZEN-BIOSCIENCE (Chengdu, Sichuan, China), while Nrf2 (16396-1-AP) and Keap1 (10503-2-AP) were form Proteintech Group (Wuhan, Hubei, China).

Techniques: Staining, Transwell Migration Assay, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: Sinensetin serves as an AMPK activator to inhibit RANKL-induced osteoclastogenesis via osteoclast cytoskeleton reorganization

doi: 10.1186/s12967-025-06708-8

Figure Lengend Snippet: SIN inhibits osteoclastogenesis associated with AMPK activity in vivo. a Histological analysis of tibia with TRAcP staining (n = 6 per group). b , c Quantitative analysis of N.Oc/BS and Oc.S/BS of TRAcP-positive cells in tibia. d Representative images of western blot showed the expression of bone proteins including CTSK and ATP6V0D2 standardized to β-actin. e , f Quantitative analysis of CTSK and ATP6V0D2 normalized to β-actin (n = 3). g , i Histological analysis of tibia with p-PAK4 and p-AMPK staining (n = 6 per group). h , j Quantitative analysis of staining area in tibia bone tissue. * p < 0.05, ** p < 0.01, *** p < 0.001. All data are expressed as mean ± SD

Article Snippet: Primary antibodies against c-Fos (ab134122), ATP6V0D2 (ab236375), p-AMPK (ab133448) and AMPK (ab32047) were obtained from Abcam (Cambridge, UK), NFATc1 (sc-7294) and CTSK (sc-48353) were from Santa Cruz Biotechnology (Dallas, USA), p-PAK4 (bs-2270R) were obtained from Bioss (Beijing, China), Cortactin ( R23963 ) were obtained from ZEN-BIOSCIENCE (Chengdu, Sichuan, China), while Nrf2 (16396-1-AP) and Keap1 (10503-2-AP) were form Proteintech Group (Wuhan, Hubei, China).

Techniques: Activity Assay, In Vivo, Staining, Western Blot, Expressing

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Red blood cell-tumor cell interactions promote tumor cell progression.

doi: 10.1186/s13046-025-03376-w

Figure Lengend Snippet: Fig. 3 Transcriptomic and functional analysis of MDA-MB-231 cells primed with RBCs or non-primed. (A) Heatmap showing results of the hierarchical clustering analysis of the significantly differentially expressed genes in MDA-MB-231 primed with RBCs from either HD (green) or M1 patients (purple) (n= 3 per group). (B) Volcano plot for the differentially expressed genes between HD and M1 samples. (C) Gene ontology analysis of the differentially expressed genes displaying the biological processes altered on MDA-MB-231 after co-cultivation with RBCs. (D) PAK4, VIM and PLS3 gene expression was analyzed by RT-qPCR, from MDA-MB-231 samples co-cultured with HD, M0 or M1 RBCs. Samples were relativized to B2M and normalized to Δct from negative control (non-primed cells, Ctrl) (n= 14 per group, triplicates). (E) A representative immunofluorescence image (left panel) highlights vimentin ex pression (in red) in MDA-MB-231 cells primed with M1 RBCs, with polarized regions marked by white arrows. Boxplot showing the percentage of polarized MDA-MB-231 cells in each group: Crtl(n=4) or after priming with RBCs from HD and M1 patients (n=6, each) (right panel). (F) Graph representing migra tion of MDA-MB-231 co-cultured with metastatic breast cancer RBCs or without RBCs in the presence or absence of PAK4 inhibitor (PAKi, LCH-7749944) (n= 5 per group, duplicates). (G) Graph representing adhesion to collagen I of MDA-MB-231 primed with metastatic breast cancer RBCs or without RBCs in the presence or absence of PAK4i (n= 5 per group, triplicates). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: PAK4 Inhibition MDA-MB-231 cells were exposed for 4 h to 10–60 μM of the PAK4 inhibitor LCH-7,749,944 (Selleckchem) to determine the minimum dose of PAK4 inhibitor that affects proliferation (n = 3/group, quadruplicates).

Techniques: Functional Assay, Gene Expression, Quantitative RT-PCR, Cell Culture, Negative Control, Immunofluorescence