Journal: Journal of Translational Medicine

Article Title: Integrative transcriptomic and experimental analysis identifies GREM1 as a pro-metastatic mediator in lung adenocarcinoma

doi: 10.1186/s12967-026-08793-9

Figure Lengend Snippet: GREM1 interacts with BMP2 and regulates TGF-β/SMAD signaling pathway. ( A ) schematic diagram illustrates protein-protein interaction networks involving hub genes and EMT, glycolysis and TGF-β signaling pathways genes, based on findings from the STRING database. ( B ) 3D Binding model analysis(GREM1 in pink and BMP2 in green). ( C ) Immunofluorescence double staining detecting the colocalization and expression changes of GREM1 (red) and BMP2 (green) in the control group, GREM1 knockdown group (sh-GREM1-1), and overexpression group (oe-GREM1). ( D ) Co-immunoprecipitation assay verifying the direct binding between GREM1 and BMP2. ( E ) Western blot analysis of TGF-β/SMAD pathway components, including SMAD2/3, p-SMAD2, p-SMAD3, TGF-β1, BMP2, Smad1, Smad5 and Smad8. ( F ) IHC staining of GREM1 in tumor and adjacent normal tissues. ( G ) Representative images of GREM1 expression in immunohistochemical staining. ( H ) Kaplan–Meier curves for overall survival by GREM1 expression

Article Snippet: The protein was separated, transferred, and then incubated with 5% milk for 2 h. Antibodies including GREM1 (Santa Cruz, sc-515877), TGF-β1 (Santa Cruz, sc-130348), smad23 (Santa Cruz, sc-398844), p-smad2 (ABclonal, AP1007), p-smad3 (ABclonal, AP0727), smad1 (ABclonal, A19113), smad5 (ABclonal, A22749), smad8 (Santa Cruz, sc-518051), BMP2(ABclonal, A27101 ), snail(ABclonal, A11794), E-cadherin (Proteintech, Cat No. 20874-1-AP), N-cadherin (proteintech, Cat No. 22018-1-AP), Vimentin (Santa Cruz, sc-373717), HK2 (Proteintech, Cat No. 22029-1-AP), LDHA (ABclonal, A21893), PKM2 (ABclonal, A0268), TPI1(ABclonal, A2579) and β-Actin (ABclonal, AC038) were incubated with the protein overnight at 4 °C.

Techniques: Protein-Protein interactions, Binding Assay, Immunofluorescence, Double Staining, Expressing, Control, Knockdown, Over Expression, Co-Immunoprecipitation Assay, Western Blot, Immunohistochemistry, Immunohistochemical staining, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Inflammation-induced osteogenic signaling promotes calcium phosphate crystal formation in kidneys via MAPK, NF-κB, and smad pathways

doi: 10.3389/fcell.2026.1831072

Figure Lengend Snippet: Cytokines Activated Their Downstream Pathways in HK-2 Cells. The phosphorylation of ERK, JNK, p38, and NF-κB in HK-2 cells treated with TNF-α (A) or IL-1β (B) was analyzed over a time course ranging from 15 min to 4 h. The phosphorylation of Smad1/5/9 and Smad2/3 in HK-2 cells treated with TGF-β1 (C) or BMP-2 (D) was analyzed over a time course ranging from 15 min to 4 h. GAPDH was served as the loading control.

Article Snippet: Primary antibodies for immunoblotting such as OPN (A19092), ALP (A0514), OPG (A2100), Runx2 (A2851), p-JNK1/2/3 (AP0631), JNK1/2/3 (A4867), p-NF-κB (AP0944), NF-κB (A19653), p-p38 (AP0057), p38 (A14401), p-ERK (AP0485), ERK (A4782), p-Smad1/5/9 (AP0850), Smad1 (A21734), p-Smad2/3 (AP0548), Smad2/3 (A7536), TNF-α (A11534), IL-1β (A11534), TGF-β1 (A16640), BMP-2 (A0231), β-actin (AC028), α-Tubulin (A6830), and GAPDH (A19056) were from ABclonal (Woburn, MA, United States).

Techniques: Phospho-proteomics, Control

Journal: Environmental Health Perspectives

Article Title: Evaluating the Effects of Neburon Exposure on Ovarian Folliculogenesis Using Zebrafish and Mouse Granulosa Cell Line

doi: 10.1021/EHP.6c00111

Figure Lengend Snippet: Expression of key genes and proteins in the Amh/Bmpr2a pathway in WT female zebrafish ovaries after 150-day exposure to neburon. (A) Transcriptional levels of amh , bone morphogenetic protein receptor 2a ( bmpr2a ), and bmpr2b in ovaries ( n = 6 samples/group) compared to control fish. (B) Protein levels of Bmpr2, phosphorylated small mother against decapentaplegic (pSmad1), and Smad1 in ovaries ( n = 3 samples/group) compared to control fish. (C) The representative ovarian sections were labeled with pSmad1 using immunostaining. Areas in brown indicated positive staining signal. The positive signal area was magnified in the lower panel, and the granulosa cells were indicated by the arrow. Scale bars, 200 μm (top panel) and 100 μm (bottom panel), respectively. (D) The proportion of pSmad1-positive (pSmad1 + ) area to total follicles area in ovarian sections ( n = 4 samples/group). (E) Protein levels of Fshr in ovaries ( n = 3 samples/group) of control and neburon-exposed fish. Data are presented as the mean ± SE. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by posthoc Duncan ’s test. Different letters on the bars represent statistically significant differences at p < 0.05 between groups. The data presented in A,B,D,E are also displayed in Excel Table S19, S20, S21 and S22 , respectively.

Article Snippet: They were then incubated overnight at 4 °C with the following primary antibodies: rabbit anti-BMPR2 primary antibody (ABclonal A5666, 1:1000), rabbit anti-pSMAD1 primary antibody (ABclonal AP0295, 1:1000), rabbit anti-SMAD1 primary antibody (ABclonal A1101, 1:1000), rabbit anti-CYP1A1 primary antibody (Proteintech 13241–1-AP, 1:1000), mouse anti-AHR primary antibody (Proteintech 67785–1–1G, 1:1000), rabbit anti-AMHR2 primary antibody (ABclonal A6523, 1:1000), rabbit anti-β-ACTIN primary antibody (ABclonal AC026, 1:4000), mouse anti-GAPDH primary antibody (Proteintech 60004–1, 1:10000) or rabbit anti-LAMIN B primary antibody (Beyotime AF1408, 1:4000).

Techniques: Expressing, Control, Labeling, Immunostaining, Staining