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l cysteine  (Thermo Fisher)
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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n acetyl l cysteine  (MedChemExpress)
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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l cysteine cys  (Tokyo Chemical Industry)
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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nac  (MedChemExpress)
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to <t>ROS/NLRP3.</t> (A) MC3T3-E1 was cultured with ROS inhibitor <t>NAC</t> (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, <t>N-acetyl-L-cysteine;</t> NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.
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H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to ROS/NLRP3. (A) MC3T3-E1 was cultured with ROS inhibitor NAC (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, N-acetyl-L-cysteine; NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.

Journal: International Journal of Molecular Medicine

Article Title: Betulinic acid alleviates the inflammatory injury of osteoblasts in osteoporosis by augmenting autophagy via the AMPK-mTOR signaling pathway

doi: 10.3892/ijmm.2026.5858

Figure Lengend Snippet: H 2 O 2 -induced osteogenic differentiation decline in MC3T3-E1 cells related to ROS/NLRP3. (A) MC3T3-E1 was cultured with ROS inhibitor NAC (20 μ M) or NLRP3 inhibitor MCC950 (100 μ M) in a medium containing H 2 O 2 (200 μ M) to produce ROS, and then intracellular ROS was detected by DCFH-DA. Scale bar, 100 μ m. (B) The average relative DCF fluorescence intensity on each pore cell was evaluated and quantified. (C) The protein levels of NLRP3 were determined by western blotting. (D) Quantification of the results shown in C. (E and F) Quantitative analysis of the expression of osteoblast marker mRNA in osteoblasts treated with NAC or MCC950 with or without H 2 O 2 by reverse transcription-quantitative PCR. (G) The protein levels of related markers in the process of osteoblast induction were determined by western blotting. (H and I) Quantification of the results shown in G. Data are expressed as the mean ± SEM. ** P<0.01 vs. control group; # P<0.05 and ## P<0.01 vs. corresponding H 2 O 2 group; & P<0.01 vs. corresponding H 2 O 2 group. ROS, reactive oxygen species; NAC, N-acetyl-L-cysteine; NLRP3, NOD-like receptor pyrin domain-containing 3; RUNX2, runt-related transcription factor 2.

Article Snippet: BA, 3-Methyladenin (3-MA), ROS scavenger N-acetyl-L-cysteine (NAC), NLRP3 inhibitor MCC950 and Dorsomorphin (synonyms: Compound c, CC) were purchased from MedChemExpress, and Essential Medium α (α-MEM) and fetal bovine serum (FBS) were purchased from Gibco, Thermo Fisher Scientific, Inc.

Techniques: Cell Culture, Fluorescence, Western Blot, Expressing, Marker, Reverse Transcription, Real-time Polymerase Chain Reaction, Control