human cmklr1  (Obio Technology Corp Ltd)

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 is overexpressed in OSCC and is associated with a poor prognosis. Relative mRNA expression of CMKLR1 in (A) HNSCC tissues ( n = 520) versus adjacent normal tissues ( n = 44) and in (B) non–lymph node (N0) versus lymph node (N1) samples, according to TCGA database analysis. (C) Kaplan–Meier survival analysis of OS based on CMKLR1 expression in patients with HNSCC, with data from TCGA. (D) Representative IHC images depicting CMKLR1 expression in OSCC tissues and adjacent normal oral mucosal tissues ( n = 68). Scale bar = 20 μm. (E) Quantitative analysis of CMKLR1 IHC staining intensity in OSCC samples ( n = 68) versus normal tissue samples, with subgroup comparisons based on T stage, clinical stage, and lymph node metastasis. (F) Kaplan–Meier survival analysis of patients with OSCC with high versus low CMKLR1 expression according to IHC scores. (G) RT-qPCR analysis of CMKLR1 mRNA expression in 50 paired OSCC and adjacent normal tissues. (H) Western blot analysis of CMKLR1 protein levels in five representative paired OSCC (T) and adjacent nontumorous (N) tissues. N, adjacent nontumorous tissues; T, OSCC tumor tissues; TCGA, The Cancer Genome Atlas. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Western Blot

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes OSCC cell proliferation and tumor growth in vivo. (A) Western blot analysis confirming CMKLR1 knockdown in Cal-27 cells (Sh1 and Sh2) and CMKLR1 overexpression in SCC-9 cells (OE). (B) CCK-8 proliferation assays of the proliferation of CMKLR1-silenced Cal-27 cells and CMKLR1-overexpressing SCC-9 cells. (C, D) Colony formation assays and quantification in the indicated groups. (E) Representative images of xenograft tumors derived from OSCC cells with CMKLR1 knockdown (Sh), OSCC cells with CMKLR1 overexpression (OE), and control cells (NC). (F) Tumor growth curves demonstrating tumor volume changes over time. (G) Tumor weight measurements at the endpoint. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: In Vivo, Western Blot, Knockdown, Over Expression, CCK-8 Assay, Derivative Assay, Control

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes OSCC cell migration, invasion, and EMT. Representative images and quantification of wound healing assays in (A) Cal-27 cells with CMKLR1 knockdown and (B) SCC-9 cells with CMKLR1 overexpression. Transwell invasion assays and quantification in (C) Cal-27 and (D) SCC-9 cells, revealing a decreased and increased invasive capacity after CMKLR1 knockdown and overexpression, respectively. Scale bar = 100 μm. (E) Western blot analysis of EMT markers (E-cadherin, N-cadherin, vimentin, Snail, and Slug) in CMKLR1-modified OSCC cells. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Migration, Knockdown, Over Expression, Western Blot, Modification

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 knockdown suppresses mitochondrial OXPHOS in OSCC cells. (A) Volcano plot of DEPs in CMKLR1-knockdown and control Cal-27 cells, depicting upregulated proteins (red), downregulated proteins (blue), and nonsignificant proteins (gray). (B) Heatmap of the top DEPs identified through proteomic analysis. (C) Subcellular localization pie chart of DEPs, revealing a large proportion localized to mitochondria. (D, E) GO and KEGG pathway enrichment analyses of DEPs, highlighting enrichment in OXPHOS-related processes. (F) Gene set enrichment analysis demonstrating the significant downregulation of OXPHOS in CMKLR1-silenced cells. (G) Seahorse XF analysis of mitochondrial respiration indicating a reduced OCR in CMKLR1-knockdown OSCC cells. (H) Quantification of basal respiration, ATP production, maximal respiration, and spare respiratory capacity in CMKLR1-knockdown cells. (I) Western blot analysis of representative OXPHOS complex subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC1, CIV-MTCO2, and CV-ATP5A1), with β-actin as loading control. (J) Representative MitoTracker images and quantification of fluorescence intensity (MitoTracker Red, scale bar = 20 μm). * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Knockdown, Control, Western Blot, Fluorescence

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes mitochondrial OXPHOS through the PI3K/AKT/PGC-1α axis. (A) Immunoblot analysis of p-PI3K, PI3K, p-AKT, AKT, and PGC-1α in OSCC cells with CMKLR1 knockdown. (B) Protein levels of p-PI3K, p-AKT, and PGC-1α were measured in OSCC cells overexpressing CMKLR1, with or without LY294002 treatment (10 μM). (C) Western blot analysis of representative OXPHOS complex subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC1, CIV-MTCO2, and CV-ATP5A1) in CMKLR1-overexpressing cells after transfection with siPGC-1α or siNC. (D) Seahorse XF analysis of OCR curves. (E) Quantification of basal respiration, ATP-linked respiration, maximal respiration, and spare respiratory capacity. (F) Representative MitoTracker images and quantification of fluorescence intensity (MitoTracker Red, scale bar = 20 μm). * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Western Blot, Knockdown, Transfection, Fluorescence

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: Inhibition of OXPHOS reverses the impact of CMKLR1-mediated tumor-promoting effects. (A) CCK-8 assay of the proliferation of CMKLR1-overexpressing (OE) OSCC cells treated with IACS-010759. (B) Colony formation assay of NC, OE, and OE + IACS-010759 cells. The right panel presents the number of colonies. (C, D) Transwell assay revealing significantly impaired cell invasion and migration after OXPHOS inhibitor treatment in CMKLR1-overexpressing cells. Scale bar = 100 μm. (E) Representative images of xenograft tumors derived from NC, OE, and OE + IACS-010759 cells. (F) Tumor growth curves demonstrating tumor volume changes over time. (G) Tumor weight measurements at the endpoint. (H) Representative images of IHC staining of tumor sections with Ki-67. Scale bar = 50 μm. (I) Quantification of Ki67 expression in xenograft tumors. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Inhibition, CCK-8 Assay, Colony Assay, Transwell Assay, Migration, Derivative Assay, Immunohistochemistry, Expressing

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: Schematic diagram of the mechanism by which CMKLR1 facilitates OSCC progression.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques:

Journal: Cells

Article Title: Simulated Microgravity Affects Pro-Resolving Properties of Primary Human Monocytes

doi: 10.3390/cells13010100

Figure Lengend Snippet: Antibodies used for immunostaining in polychromatic flow cytometry experiments.

Article Snippet: ChemR23 , APC , Miltenyi Biotec, Bergisch Gladbach, Germany , 1:50.

Techniques: Immunostaining, Flow Cytometry

Journal: Cells

Article Title: Simulated Microgravity Affects Pro-Resolving Properties of Primary Human Monocytes

doi: 10.3390/cells13010100

Figure Lengend Snippet: Protein expression of SPM receptors and metabolic enzymes in lymphocytes and monocytes cultured for 24 h in RCCS-simulated microgravity. ( A ) Representative scatter plot showing the gating strategy for polychromatic flow cytometry experiments: T lymphocytes and monocytes were gated as CD3+ and CD14+ cells, respectively, after being appropriately gated according to their viability and physical parameters (1 × 10 6 cells per condition), before measuring the immunoreactivity of each marker. ( B – E ) Expression of GPR32, ALX, GPR18, ChemR23 and 5-LOX in CD3+ T lymphocytes ( B , C ) and CD14+ monocytes ( D , E ). Data are shown as arbitrary units of mean fluorescence intensity (MFI) of six to nine independent experiments. * p < 0.05 (paired Student’s t -test). ( F , G ) Representative plots showing the distribution of immunofluorescence (polychromatic flow cytometry) for GPR32 and GPR 18 ( F ) and 5-LOX ( G ) in CD14+ cultured at 1 g- and μg.

Article Snippet: ChemR23 , APC , Miltenyi Biotec, Bergisch Gladbach, Germany , 1:50.

Techniques: Expressing, Cell Culture, Flow Cytometry, Marker, Fluorescence, Immunofluorescence

Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: Primer sequences used for qPCR .

Article Snippet: The point mutant plasmid of human CMKLR1 was purchased from Shanghai Generay Biotechnology.

Techniques:

Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: CMKLR1 signaling is associated with fat thermogenesis and fatness. (a–e) CMKLR1 screening as a potential GPCR associated with fat thermogenesis via transcriptomic analysis. Brown adipose tissue and subcutaneous WAT were dissected from mice that were treated in thermo-neutral (30 °C) and cold (4 °C) temperatures for 72 h. A total of 12 samples with three replicates for each condition were evaluated. (a) Flowchart of screening. (b) Four groups DEGs from transcriptome were analyzed by using a Venn diagram. iWAT-TN-vs-BAT-TN Up-regulated: the up-regulated DEGs in the thermal-neutral iWAT group compared with thermal-neutral BAT group. iWAT_TN-vs-iWAT_C: the DEGs between the thermal-neutral iWAT group and cold temperature iWAT group. iWAT_C-vs-BAT_C Down-Regulated: the down-regulated DEGs in the cold temperature iWAT group compared with cold temperature BAT group. BAT_TN-vs-BAT_C: the DEGs between thermal-neutral BAT group and cold temperature BAT group. (c, d) The RPKM of 19 GPCR genes in mouse fat (c) from Mouse ENCODE transcriptome data (PRJNA66167) and human fat (d) from HPA RNA-seq normal tissues (PRJEB4337). (e) The TPM of ADRB3 and CMKLR1 in human fat from GTEx database. (f, h) The level of chemerin (f) and RvE1 (h) in the serum of lean and overweight/obese people. Two ELISA kits were used to measure the level of chemerin and RvE1. Lean (BMI = 18.5∼23.9) n = 21, Overweight/Obese (BMI > 25.5) n = 21. (g) Correlation between the serum chemerin level and body mass index (BMI). (i) Correlation between the serum RvE1 level and body mass index (BMI). (j–k) The serum level of chemerin (j) and RvE1 (k) was measured in the serum of mice fed either a NCD or HFD. Two ELISA kits were used to measure the level of mouse chemerin and RvE1. TN, thermo-neutral; C, cold temperature; iWAT, inguinal white adipose tissue; BAT, brown adipose tissue; RPKM, Reads Per Kilobase per Million mapped reads; TPM, Transcripts Per Kilobase Million; GPCR, G-protein-coupled receptor; BMI, body mass index; NCD, normal chow diet; HFD, high-fat diet; RvE1, Resolvin E1. All data are presented as mean ± SEM . Statistical significance was determined by unpaired two-tailed student's t-test (f, h and j, k) or simple linear regression (g, i).

Article Snippet: The point mutant plasmid of human CMKLR1 was purchased from Shanghai Generay Biotechnology.

Techniques: RNA Sequencing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: Gene annotation for screened genes. The 420 screened genes were annotated by David database and 20 of these genes were identified as G-protein coupled receptor encoding gene.

Article Snippet: The point mutant plasmid of human CMKLR1 was purchased from Shanghai Generay Biotechnology.

Techniques: Activity Assay

Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: Chemerin and RvE1 differentially regulated mTORC1 signaling via CMKLR1. (a) Gene set enrichment analysis (GSEA) analysis for gene signatures of mTOR signaling pathway in iWAT from Rarres2 KO compared with C57BL/6J control mice. NES, normalized enrichment score. FDR, false discovery rate. (b, c) Western-blot analysis for level of p-S6K and S6K protein in undifferentiated 3T3-L1 treated with 100 ng/ml chemerin (b) or 100 ng/ml RvE1 (c). (d, e) Western-blot analysis for level of UCP1, p-S6K and S6K protein in 3T3-L1 mature adipocytes treated with 1 µM CL, 1 µM CL with 100 ng/ml chemerin (CL+CHE), 10 µM Mi or 100 ng/ml (0.285 µM) RvE1. (f) C57BL/6J mice fed with NCD for 8 weeks were injected with Vehicle, chemerin-9 (1 µg/mouse/day), Rapamycin (2 mg/kg/day) or Rapamycin (2 mg/kg/day) with chemerin-9 (1 µg/mouse/day) over 7 days. Western-blot analysis for level of UCP1 protein in iWAT. (g) C57BL/6J mice fed with NCD for 8 weeks were injected with Vehicle, RvE1 (1 µg/mouse/2day), Rapamycin (2 mg/kg/2day) or Rapamycin (2 mg/kg/2day) with RvE1 (1 µg/mouse/2day) over 14 days. Western-blot analysis for level of UCP1 protein in iWAT. (h-i) Western-blot analysis for level of p-S6K, S6K, UCP1 and CMKLR1 protein in different 3T3-L1 cell pools treated with 100 ng/ml chemerin, 10 µM Mirabegron or 100 ng/ml RvE1 as well as transfected with Cas9 and guide RNA targeting the Cmklr1 gene sequence. gNS indicates 3T3-L1 cell pools transfected with Cas9 only and g#1 indicates 3T3-L1 cell pools transfected with Cas9 and guide RNA. The ImageJ software was used for gray scanning.

Article Snippet: The point mutant plasmid of human CMKLR1 was purchased from Shanghai Generay Biotechnology.

Techniques: Control, Western Blot, Injection, Transfection, Sequencing, Software

Journal: Fundamental Research

Article Title: CMKLR1 senses chemerin/resolvin E1 to control adipose thermogenesis and modulate metabolic homeostasis

doi: 10.1016/j.fmre.2022.06.014

Figure Lengend Snippet: RvE1 and chemerin exert differential regulatory effects by forming hydrogen bonds with different binding sites of CMKLR1. (a, b) The binding modes of RvE1 (a) and chemerin-9 (b) to human CMKLR1 (hCMKLR1). hCMKLR1 is shown as a slate cartoon model, RvE1 and chemerin-9 are shown as a cyan sticks model, amino acid residues corresponding to each amino acid binding site are highlighted in forest and hydrogen bonds formed between the ligand and each amino acid residue are indicated by yellow dotted lines. The hydrogen bond distances are listed next to the red dotted line. Hydrogen bond donor atoms are shown in red and hydrogen bond acceptor atoms are shown in blue. (c) qPCR analysis of CMKLR1 in Hela. All data are presented as mean ± SEM . Statistical significance was determined by unpaired two-tailed Student's t-test. (d–f) Western-blot analysis for level of CMKLR1, p-S6K and S6K protein in Hela treated with 1 µM chemerin-9 or 100 ng/ml RvE1 as well as transfected with different plasmids of human CMKLR1. The ImageJ software was used for gray scanning.

Article Snippet: The point mutant plasmid of human CMKLR1 was purchased from Shanghai Generay Biotechnology.

Techniques: Binding Assay, Residue, Two Tailed Test, Western Blot, Transfection, Software