Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: Adapalene reduces LPS-stimulated pro-inflammatory cytokine production in murine macrophages. (A) RAW264.7 cells were treated with adapalene (10, 100, or 1000 nM) for 1 h, followed by LPS (100 ng/mL) for 24 h. Total RNA was prepared for qRT-PCR analysis of TNFα, IL-1β, IL-6, iNOS, and COX-2, and mRNA levels were determined using gene-specific primers. (B) Following pretreatment with or without adapalene, TNFα, IL-6, and IL-1β levels in culture media were quantified using ELISA. NO and PGE2 levels in culture media were quantified using a Griess reaction assay and EIAs, respectively. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. LPS alone. (C) mRNA expression of M2 markers MRC1 and Arg1 was determined using RT-qPCR. (D) Protein levels of pro-inflammatory cytokines and M2 markers were determined using western blot analysis. Band densities were analyzed using ImageJ software, normalized to β-actin, and the results are shown in lower panel. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05 and ## P < 0.01 vs. LPS alone. (E, F) After treatment of RAW264.7 cells with LPS and adapalene, immunofluorescence analysis of F4/80 positive remove ( + )macrophages was carried out. Flow cytometry data were analyzed using FlowJo Software v11.0 (Ashland, OR, USA). The experiment was performed three times in triplicate and results are presented as the mean ± S.D. of three independent experiments. A complete flow cytometry gating strategy and isotype control data are shown in Supplementary Fig. 1E. AD; adapalene.

Article Snippet: COX-2 , Santa Cruz , sc-376,861 , Mouse , 1:500.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Software, Incubation, Control, Immunofluorescence, Flow Cytometry

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: In vivo anti-inflammatory effects of adapalene in LPS-induced septic shock model. Six mice per group were treated with vehicle only or adapalene (20, 50, or 100 mg/kg, p.o. ) for 1 h and then injected with LPS (25 mg/kg, i.p. ). Serum and liver samples were collected from each mouse after 6 h. (A, B) TNFα, IL-6, and IL-1β protein in serum (A) and mRNA levels of TNFα, IL-1β, IL-6, iNOS, and COX-2 in liver (B) were determined using EIA and qRT-PCR, respectively. (C) TNFα, IL-1β, IL-6, iNOS, COX-2, MRC1, Arg1, RARα, RARβ, and RARγ protein levels in liver were determined using western blot. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Density analysis is shown in the right panel. (D) Protein levels of p38, p-p38 MAPK, ERK, p-ERK, JNK, p-JNK, PI3K, p-PI3K, Akt, p-Akt, STAT3, and p-STAT3 in the liver were determined using western blotting. Density analysis is shown in the lower panel. Results are presented as the means ± S.D. of six mice. Representative images of p-NF-κB staining (E), F4/80 staining (F), H&E staining (H), and Picro-Sirius Red staining (K) of liver tissues were shown. Quantitative analysis of p-NF-κB and F4/80 staining was performed using ImageJ software (G). The survival rate after adapalene administration in LPS-induced sepsis is shown (I). Hepatic mRNA and protein levels of fibrosis markers were determined (J). AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. LPS alone.

Article Snippet: COX-2 , Santa Cruz , sc-376,861 , Mouse , 1:500.

Techniques: In Vivo, Injection, Quantitative RT-PCR, Western Blot, Incubation, Staining, Software, Control

Journal: Scientific Reports

Article Title: Adapalene, an RAR agonist, exerts anti-inflammatory effects by regulating macrophage polarization through RAR -mediated signaling pathways

doi: 10.1038/s41598-026-44454-z

Figure Lengend Snippet: In vivo anti-inflammatory effects of adapalene in HFD-induced obesity model. Adapalene (10 or 50 mg/kg) was orally administered once daily to HFD-induced obese mice ( n = 10 per group) for 3 weeks. On the last day of administration, serum and liver samples were collected from each mouse. (A) TNFα, IL-1β, IL-6, iNOS, COX-2, MRC1, Arg1, p38, p-p38 MAPK, ERK, p-ERK, JNK, p-JNK, RARα, RARβ, and RARγ protein levels in the liver were determined using western blotting. Density analysis is shown in the right panel. (B) PI3K, p-PI3K, Akt, p-Akt, STAT3, and p-STAT3 protein levels in the liver were determined using western blotting. Membranes were cut prior to antibody incubation; all available uncropped images are provided in the Supplementary Information. Original full-length images for certain replicates are unavailable due to loss of original acquisition files. Density analysis is shown in the lower panel. (C) TNFα, IL-6, IL-1β, COX-2, iNOS, MRC1, and Arg1 mRNA levels in the liver were determined using qRT-PCR. (D) Representative images of Oil Red O staining and Picro-Sirius Red staining (G) of liver tissues are shown. Results are presented as the means ± S.D. of eight mice. (E, F) Hepatic protein levels (E), mRNA levels of fibrosis markers (F) and serum ALT and AST levels (F) were determined. AD; adapalene, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control; # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. LPS alone.

Article Snippet: COX-2 , Santa Cruz , sc-376,861 , Mouse , 1:500.

Techniques: In Vivo, Western Blot, Incubation, Quantitative RT-PCR, Staining, Control