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: High Calcium and Pro-inflammatory Cytokines Increased the Levels of Osteogenesis-Related Proteins in HK-2 Cells. Western blot analysis of ALP, OPN, and OPG in HK-2 cells treated with CaCl 2 (A) TNF-α (B) IL-1β (C) TGF-β1 (D) or BMP-2 (E) for 6 h. The quantitative results showed the fold change of the protein expression compared to the control group. β-actin or tubulin served as the loading control. Data are presented as individual data points (scatter plot) with the mean ± SEM; n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001 versus the control group (One-way ANOVA).

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: Western Blot, Expressing, Control

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: Pro-inflammatory Cytokines Upregulated Osteogenesis-related Proteins via the MAPK/NF-κB and Smad Signaling Pathway. HK-2 cells were pretreated with 10 µM ERK inhibitor (U0126), JNK inhibitor (SP600125), p38 inhibitor (SB203580), or NF-κB inhibitor (BAY 11–7,082) for 2 h, and then treated with 20 ng/mL TNF-α (A) or IL-1β (B) for 72 h (C) HK-2 cells were pretreated with 5 µM Smad2/3 inhibitor (SB-525334) for 2 h and then treated with 200 ng/mL TGF-β1 for 72 h (D) HK-2 cells were pretreated with 10 µM Smad1/5/8 inhibitor (LDN-193189) for 2 h and then treated with 100 ng/mL BMP-2 for 72 h. The expression of osteoblast markers (ALP, OPN, and OPG) was examined by Western blotting. The quantitative results showed the fold change of the protein expression compared to the control group. GAPDH served as the loading control. Data are presented as individual data points (scatter plot) with the mean ± SEM; n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001 versus the control group; # P < 0.05, ## P < 0.01, ### P < 0.001 versus cytokine group (One-way ANOVA).

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: Expressing, Western Blot, Control

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: Osteogenesis-related Proteins Directly Regulate Crystal Nucleation, Growth, and Aggregation. (A) CaP crystallization in the presence of OPG, ALP, or OPN was quantified by alizarin red staining. Alizarin red was extracted from the stained crystal and the absorbance was read at 405 nm. The quantitative results showed the fold change of the absorbance compared to the control group. (B) CaP crystals were incubated with OPG, ALP, or OPN for 60 min, the change in the crystal size indicated the crystal growth. The quantitative results indicated the fold change in crystal growth of the experimental group compared to that of the control group. (C) CaP crystals were stirred with ALP, OPN or OPG for 60 min, the number and area of the aggregates were counted. The average area per aggregate was calculated by the formula: The total area of the aggregates in each field/the number of the aggregates in each field. The quantitative results showed the fold change to the control group. Data are mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 versus the control group (one-way ANOVA, n = 3).

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: Crystallization Assay, Staining, Control, Incubation

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: Representative Immunohistochemical Staining of Inflammatory Cytokines and Osteoblast Markers in the Kidneys of Rats Fed a High-Calcium Diet. Representative images of TNF-α, IL-1β, TGF-β1, BMP-2 (inflammatory cytokines) and OPN, OPG, ALP, Runx2 (osteogenesis-related proteins) in kidney tissues from each experimental group are shown (400×). IHC staining was performed to visualize the spatial distribution of target proteins within renal tissue. Scale bar, 20 µm.

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: Immunohistochemical staining, Staining, Immunohistochemistry

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: Hypothesized Model of Randall’s Plaque Formation. a, In a high-calcium renal environment, activated macrophages and inflammasomes release pro-inflammatory cytokines, activating signaling pathways such as MAPK, NF-κB, and Smad in renal tubular cells. b, These signaling pathways upregulate proteins involved in osteogenesis, such as Runx2, ALP, OPG, and OPN, and result in an osteogenic transition. c, Calcium-enriched and osteogenesis-related protein-enriched vesicles are released from the basal side of tubule epithelial cells. Osteogenesis-related proteins directly promote the nucleation, growth and aggregation of CaP crystals in these vesicles. d, CaP aggregates-enriched vesicles move to the renal interstitium, calcifying the interstitial collagen, and eventually leading to the formation of Randall’s plaque in the renal papillae. This figure was created by BioRender.com .

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: Protein-Protein interactions

Journal: NPJ Precision Oncology

Article Title: Multi-omics profiling reveals tumor microenvironment characteristics linked to immunotherapy response and prognosis in non-small cell lung cancer

doi: 10.1038/s41698-026-01474-2

Figure Lengend Snippet: A Identification of four celltypes in the LLC TME by scRNA-seq. B Anti-SPP1 therapy increases all immune subsets, decreases non-immune cells. C Subclustering of T/NK cells. D GO enrichment analysis of genes highly expressed in CD8 + T cells at different groups (control vs. anti-SPP1). E Anti-SPP1 therapy decreases M2-like macrophage proportion. F Immunofluorescence indicated increased CD8 + T infiltration with concurrent M2 reduction post-treatment.

Article Snippet: Five days post-inoculation, mice with palpable tumors were randomly assigned to four treatment groups ( n = 4 per group) using a random number table : Control (PBS) , Anti-PD-1 (clone RMP1-14, Bio X Cell, #BE0146) , Anti-SPP1 (anti-mouse osteopontin antibody, clone 103D6, MCE), and Anti-SPP1 combined with Anti-PD-1.

Techniques: Control, Immunofluorescence