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: 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