Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: p38γ inhibitors increase olfactory transduction pathway activity. (A) WB analysis of Hut78 cells treated with 0.5 to 10 μM of CSH18. (B) WB analysis of Hut78 cells treated with 0.5 to 10 μM of CSH71. Both CSH18 and CSH71 increased OR4N5 protein levels. Other olfactory factors OR10K2, OR2H2, and OR4K17 were also assessed, with dimerization of OR4N5 observed based on molecular weight (blue arrows indicate 50 kDa). (C) WB assays for lentiviral expression of sh-p38γ vs sh-Ctrl. OR4N5 bands at ∼30 and 60 kDa correspond to the monomer and dimer, respectively (sh-Ctrl lane). (D) OR4N5 3D structure prediction by AlphaFold (AF-Q8IXE1-F1, left). Right: localization of OR4N5 in the plasma membrane (green, confidence score = 5). Slight presence in the cytoskeleton (confidence score = 1) according to Genecard.com . (E) In-depth analysis of the OR4N5 amino acid sequence identified 7 transmembrane domains (underlined) and 2 cysteines that may contribute to OR4N5 dimerization. CDR, Complementarity Determining Region; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hr, hour; sh-Ctrl, shRNAs targeting control; sh-p38γ, shRNAs targeting p38γ; TM, Transmembrane.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Transduction, Activity Assay, Molecular Weight, Expressing, Clinical Proteomics, Membrane, Sequencing, Control

Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: Gene silencing of either OR4N5 or OR10K2 promotes cell proliferation in Hut78 cells. (A) Cell viability assays were conducted on OR4N5- and OR10K2-silenced cells (sh- OR4N5 and sh- OR10K2 ) over a 10-day period after transduction to assess cell proliferation. Cell counts were measured on days 1 through 10 using the trypan blue exclusion method to distinguish viable cells. A scramble sequence shRNA served as the control in these assays. (B) Confocal immunofluorescence analysis confirmed the successful silencing of OR4N5 and OR10K2 proteins in Hut78 cells (left). The loss of OR expression in Hut78 cells correlates with increased cell proliferation 7 to 9 days after transduction. Statistical data are presented as mean (standard deviation). Cells were counted on days 1, 4, 7, and 10 after selection using an automated cell counter, with trypan blue exclusion to differentiate viable cells. sh-Ctrl, short hairpin control.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Transduction, Sequencing, shRNA, Control, Immunofluorescence, Expressing, Standard Deviation, Selection

Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: Colocalization of OR4N5 and CD3E, and colocalization of p38γ with CD3Z in TCR complexes using confocal immunofluorescence microscopy. (A) Colocalization of OR4N5 and CD3E in PBMCs from healthy donors. (B) Colocalization of OR4N5 and CD3E in CTCL Hut78 cells, both treated with CSH71 (250 nM) and untreated controls. In both panels A and B, the interaction between CD3E and OR4N5 was assessed, with colocalization indicated by the yellow signal in merged 2-color images.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Immunofluorescence, Microscopy