afatinib treatment  (MedChemExpress)

Journal: bioRxiv

Article Title: NF1 Loss Remodels Tumor Niches for Immune Evasion

doi: 10.64898/2026.01.11.698818

Figure Lengend Snippet: (A) GSEA Hallmark pathway analysis showing EMT, TGFβ, angiogenesis, and hypoxia are upregulated as a result of NF1 knockdown (B). Heatmap showing differentially expressed genes between shNF1 and shSCR expressing cells. (C) Western blots showing NF1 and EGFR expression before and after DOX-induced NF1 expression (rNF1) compared to control (rControl) in NF1 Mut STC-08-175 and Mewo melanoma cells. HSP90 served as a loading control. (D) GSEA Hallmark pathway analysis showing IFNα and IFNɣ responses are upregulated, and EMT, hypoxia and inflammatory response are downregulated as a result of NF1 ectopic expression. (E) Heatmap showing significant differential expressed genes comparing NF1 ectopic expression to control. (F) Growth curves of NF1 Mut STC-08-175 melanoma cells ectopically expressing rNF1 compared to control. P -values were calculated with a two-sided t-test at the experimental endpoint. Data points represent mean ± SD. (n=12). (G) Western blots of NF1 co-immunoprecipitation showing EGFR levels after pulling down with NF1 or IgG control antibodies after DOX-induced NF1 expression (rpbNF1) or control (rbpControl) in STC-08-175 or Mewo NF1 Mut melanoma cells. (H) Western blots comparing NF1 and EGFR, expression in shNf1 compared to shSCR RIM-3 murine melanoma cells. HSP90 served as a loading control. (I) RT-PCR data showing Nf1 and Egfr expression changes in shNf1compared to shSCR expressing RIM3 melanoma cells. Bar graphs indicate mean ± SD (n=3). P -values were calculated with a two-sided t-test. (J) Growth curves of RIM-3 melanoma cells expressing shNf1 compared to shSCR (control). P -values were calculated with a two-sided t-test at the experimental endpoint. Data points represent mean ± SD. (n=12). (K) Heatmap showing differentially expressed antigen presentation genes in RIM-3-shNF1 melanoma cells after EGFR knockdown. (L) GSEA Hallmark Gene Sets analysis showing enriched pathways in shEGFR RIM-3-shNF1 cell lines. (M) Heatmap showing differentially expressed HLA genes in shEGFR and shSCR (control) expressing NF1 Mut STC-07-127 melanoma cells. (N) Western blots comparing HLA and EGFR expression with and without afatinib treatment in three human NF1 Mut melanoma cell lines. HSP90 served as a loading control. (O) Flow cytometry analysis showing MHC class I antigen presentation difference between IFNɣ-stimulated or non-stimulated NF1 Mut STC-08-175 melanoma cells with and without afatinib treatment. Bar graphs indicate mean ± SD (n=3). P -values were calculated with a two-sided t-test.

Article Snippet: Tumor volumes were calculated with the formula: (v (π/6*l*w2), where l = length in mm, w = width in mm, v=mm3). shNF1 RIM-3 tumors were allowed to reach ∼100-200 mm 3 for growth curves or ∼500 mm 3 for flow analysis before being randomly assigned to either afatinib treatment (Medchem Express, BIBW 2992, Cat# HY-10261), PD-1(CD279) anti-mouse Mab (Bioxcell, Cat# BE0146), a combination of both treatments or the appropriate vehicle control.

Techniques: Knockdown, Expressing, Western Blot, Control, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Immunopeptidomics, Flow Cytometry

Journal: Biology Open

Article Title: Development of an adenosquamous carcinoma histopathology – selective lung metastasis model

doi: 10.1242/bio.059623

Figure Lengend Snippet: ERBB inhibition with afatinib and erlotinib decreases the number of ERBB and pERK positive cells and pan-ERBB inhibition increases the survival of mice. (A) Three representative H&E stainings of mice treated for four weeks 10 days after ASC tumor cell i.v. transplantation with either vehicle control or afatinib (AF). Scale bar: 5000 μm. n =4 (VEH); n =3 (AF). (B) Quantification of the tumor burden after treatments presented in (A). The graph represents mean ±s.d. for each group, and each point represents an individual mouse. Two-tailed unpaired Student's t -test value is *** P <0.001. n =4 (VEH); n =3 (AF). (C) The survival percentage of mice treated with vehicle, ER and AF 10 days after ASC tumor cell i.v. transplantation. Graph represents the mean ±s.d. for each group, and each point represents an individual mouse. Gehan-Breslow-Wilcoxon test values are * P <0.05. n =11 (VEH); n =8 (ER/AF). (D) Representative H&E, pAKT and pERK1/2 IHC images of source and i.v. tumors from C57BL/6 recipients upon transplantation with ASC tumor derived cells and treated with vehicle, EGFR inhibitor erlotinib (ER) or pan-ERBB inhibitor afatinib (AF). Scale bar: 500 µm (2×) or 20 µm (40×). n =5. (E-F) Quantification of pAKT and pERK1/2 positive area by dividing the positive tumor area with the total tumor area in each tumor separately by using FIJI ImageJ. The graph represents mean ±s.d. for each group, and each point represents an individual tumor. Two-tailed unpaired Student's t -test values are ** P <0.01. n =5. (G) Representative H&E, pEGFR, pERBB2 and pERBB3 IHC images of source and i.v. tumors from C57BL6 recipients upon transplantation with ASC tumor derived cells and treated with vehicle, EGFR inhibitor erlotinib (ER) or pan-ERBB inhibitor afatinib (AF) and euthanized when mice developed health problems. Scale bar: 500 µm (2×) or 20 µm (40×). n =5. (H-J) Quantification of pEGFR, pERBB2 and pERBB3 positive area by dividing the positive tumor area with the total tumor area in each tumor separately by using FIJI ImageJ. The graph represents mean ±s.d. for each group, and each point represents an individual tumor. Two-tailed unpaired Student's t -test values are * P <0.05, ** P <0.01, *** P <0.001. n =5.

Article Snippet: Treatment with afatinib (12.5 mg/kg; MedChemExpress) or erlotinib (12.5 mg/kg; MedChemExpress) in 0.5% hydroxyl propyl methyl cellulose and 0.1% Tween 80 in H 2 O by oral gavage was started 10 days after tumor cell injection.

Techniques: Inhibition, Transplantation Assay, Control, Two Tailed Test, Derivative Assay