bx 795  (InvivoGen)

Journal: bioRxiv

Article Title: A human immune system mouse model for preclinical evaluation of therapies in pemphigoid disease

doi: 10.64898/2026.01.15.699625

Figure Lengend Snippet: ( A ) Flow cytometric analysis of cytosolic ROS production in human peripheral blood cells after stimulation with COL7 c -IgG immune complexes (IC) upon pre-treatment with 1µM BX-795 or vehicle control. n=4 independent experiments using cells from different donors were performed. ( B ) Quantification of secreted human cytokines in the supernatants of COL7 c -IC stimulated human cells, comparing presence or absence of PDK1 kinase inhibitor BX-795, measured using the LEGENDplex TM Human Inflammation Panel 1. ( C ) Induction of dermal-epidermal separation in cryosections of human skin was performed upon treatment of neutrophils with 1µM BX-795 compared to untreated control cells. Line symbols the dermal-epidermal junction, asterisks separation. Scale bar 500 µm. Statistical analyses employed the Shapiro-Wilk normality test. Depending on Gaussian distribution either paired t-test or Wilcoxon test was used. * p < 0.05; ** p < 0.01.

Article Snippet: For evaluating the kinase PDK1 as a therapeutic target, HIS mice were treated orally with the small-molecule inhibitor BX-795 (Selleckchem) at 30 mg/kg daily.

Techniques: Control

Journal: bioRxiv

Article Title: A human immune system mouse model for preclinical evaluation of therapies in pemphigoid disease

doi: 10.64898/2026.01.15.699625

Figure Lengend Snippet: ( A ) Schematic representation of the experimental approach for inducing an EBA-like phenotype in human immune system mice, combined with p.o. administration of 30 mg/kg BX-795 or vehicle as control. ( B ) Assessment of skin disease severity shown as % ABSA over time (left) and as AUC (right). ( C ) Histological analysis of skin sections using PAS staining for tissue structure and infiltrate evaluation (arrow) and quantification of epidermal thickness (in µm, right). Veh. Ctrl., vehicle control. ( D ) Human immune cell composition in digested skin samples (left, per gram skin and 100000 live cells), including expression of activation markers CD86 and CD18 (middle), and intracellular ROS production (right). C., classical. ΔMFI, median fluorescence intensity after subtraction of fluorescence minus one. ( E ) Quantification of human cytokines in plasma on day 6 using the LEGENDplex TM Human Inflammation Panel 1. Symbols indicate individual mice and bars indicate means. Statistical analyses were performed using Shapiro-Wilk normality test. Depending on Gaussian distribution, samples were analyzed using unpaired t-test or Mann-Whitney test, respectively. *p < 0.05; **p < 0.01; ***p < 0.001; ****p <0.0001.

Article Snippet: For evaluating the kinase PDK1 as a therapeutic target, HIS mice were treated orally with the small-molecule inhibitor BX-795 (Selleckchem) at 30 mg/kg daily.

Techniques: Control, Staining, Expressing, Activation Assay, Fluorescence, Clinical Proteomics, MANN-WHITNEY

Journal: bioRxiv

Article Title: PDK1 and YAP1/TEAD signaling drive acquired KRAS inhibitor resistance in KRAS-mutant non-small cell lung cancer

doi: 10.64898/2025.12.22.696060

Figure Lengend Snippet: PDK1 expression is necessary and sufficient for generating resistance to KRAS G12D inhibition. (A) Growth inhibition assays of 344SQ-G12D cells treated with MRTX1133 (blue line) or co-treated with BX795 at its IC50 concentration (red line). IC50 values for MRTX1133 are indicated on the graph. (B) Growth inhibition assays of 344SQ-G12D MRTX1133-resistant line (M1133Res) treated with MRTX1133 (blue line) or co-treated with BX795 at its IC50 concentration (red line). (C) Western blot analysis of phosphorylated and total PDK1, S6, and RSK2 in 344SQ-G12D and M1133Res cells upon BX795 treatment at IC50 concentrations (3uM and 8uM, respectively). Actin was used as a loading control. (D) Colony formation assays of 344SQ-G12D and M1133Res cells upon treatment with vehicle, MRTX1133, BX795, or both. (E) Graph showing quantification of the data in D. *, p<0.05. **, p<0.01. (F) Western blot of 344SQ-G12D, M1133Res (parental control), and two PDK1 knockout clones (PDK1 KO-1 and -2) of M1133Res cells to evaluate phosphorylated and total PDK1, S6, and RSK2 levels. (G) Growth inhibition assays of M1133Res (blue), PDK1 KO-1 (red) and PDK1 KO-2 (grey) cells upon treatment with MRTX1133. (H) Colony formation assays of 344SQ-G12D, M1133Res, and PDK1 KO-1 and KO-2 cells when treated with vehicle or MRTX1133 at the IC30 concentration of the parental control line for 72 hrs. (I) Quantification of the colony formation assay in H. **, p<0.01. (J) Western blot of 344SQ-G12D cells transduced to stably express PDK1 (344SQ-PDK1) under a doxycycline-inducible promoter. The cells were treated with dox for the indicated time points before collection. The blots were probed for phosphorylated and total PDK1, S6, and RSK2 levels. (K) Growth inhibition assays of 344SQ-PDK1 cells with (blue line) or without (red line) 48 hours of dox pretreatment. The IC50 of MRTX1133 for each cell line is indicated on the graph. (L) Colony formation assays of 344SQ-PDK1 cells with or without 24 hours of dox treatment. The cells were treated with either DMSO or MRTX1133 at the IC30 concentration of 344SQ-PDK1 +dox cells. The colonies were then allowed to grow out for 3 days in drug-free complete media after which they were stained. (M) Quantification of the data in L. *, p<0.05, **, p<0.01.

Article Snippet: BX795 (Selleck Chemicals), VT107 and GSK2334470 (MedChemExpress) were dissolved in a PEG-based solvent according to the manufacturer’s instructions.

Techniques: Expressing, Inhibition, Concentration Assay, Western Blot, Control, Knock-Out, Clone Assay, Colony Assay, Stable Transfection, Staining

Journal: bioRxiv

Article Title: PDK1 and YAP1/TEAD signaling drive acquired KRAS inhibitor resistance in KRAS-mutant non-small cell lung cancer

doi: 10.64898/2025.12.22.696060

Figure Lengend Snippet: Combination treatment with G12Di and PDK1i results in tumor growth inhibition in vivo . (A) Mice implanted with 344SQ-G12D (left panel) or M1133Res cells (right panel) were treated with either vehicle (blue line), MRTX1133 (red line), BX795 (green line), or MRTX1133 and BX795 (purple line) for approximately 3 weeks. Statistical comparisons are shown for 344SQ-G12D tumors treated with vehicle vs MRTX1133 (blue asterisks) and M1133Res tumors treated with MRTX1133 vs dual combination (red asterisks) at the indicated time points. **, p<0.01. (B) IHC staining is shown for pERK (T202/Y204), Ki67, and pPDK1 (S241) levels in representative sections of tumors harvested from the mice at endpoint. Scale bars correspond to 20 uM. (C) Mice implanted with 344SQ-G12D, M1133Res, or M1133Res-PDK1-KO-2 cells. Vehicle-treated tumor growth was recorded for 344SQ (red), M1133Res (blue), and M1133Res-PDK1-KO-2 (grey). MRTX1133-treated tumor growth was recorded for 344SQ (purple), M1133Res (green), and M1133Res-PDK1-KO (orange). Treatment was continued for 4 weeks. Statistical comparisons are shown for M1133Res and M1133Res-PDK1-KO1 cohorts when treated with vehicle (blue asterisks) or MRTX1133 (brown asterisks) at the indicated time points. *, p<0.05. **, p<0.01. (D) Percent change in tumor volumes of 344SQ-G12D, M1133Res, and M1133Res-PDK1-KO-2 cohorts from at endpoint. Statistical significance was calculated using Student’s t-test. *, p<0.05. **, p<0.01. (E) IHC staining is shown for pERK (T202/Y204), Ki67, pPDK1 (S241), and pRSK2 (S227) in representative sections of tumors harvested from mice at endpoint in C. Scale bars correspond to 20uM.

Article Snippet: BX795 (Selleck Chemicals), VT107 and GSK2334470 (MedChemExpress) were dissolved in a PEG-based solvent according to the manufacturer’s instructions.

Techniques: Inhibition, In Vivo, Immunohistochemistry