Journal: Translational Lung Cancer Research

Article Title: PKC-iota drives EGFR-TKI resistance in EGFR-mutated NSCLC by phosphorylating FASN to reprogram lipid metabolism

doi: 10.21037/tlcr-2025-aw-1260

Figure Lengend Snippet: PKC-iota induces fatty acid metabolism reprogramming to increase EGFR membrane-localization via FASN. (A-E) H1975 cells were cotransfected with siRNA-FASN or siRNA-NC and Flag-PKC-iota or empty vector (worked as a control) for 48 hours: (A,B) FAs metabolites (A. SFAs; B. UFAs) of H1975 cells were analyzed by GC-MS, and normalized by total protein levels; (C) TG levels of H1975 cells were quantitated by a colorimetric method and normalized by total protein levels and normalized to 1 by the control group. ns, not significant; *, P<0.05. (D) Representative Oil Red O staining images in H1975 cells (scale bar, 100 µm); (E) representative Nile Red staining images in H1975 cells (scale bar, 75 µm). (F) After 24 hours of transfection (siRNA-FASN or siRNA-NC and Flag-PKC-iota or empty vector), H1975 cells were plated onto 3.5 cm confocal dishes for 24 hours, followed by 30 min TMA-DPH (2 µmol/L) treatment, measured fluorescence polarization and calculated membrane LFU. ns, nonspecific; *, P<0.05; ***, P<0.001. (G) IF staining was done after 48 hours of transfection (siRNA-FASN or siRNA-NC and Flag-PKC-iota or empty vector) and followed by 24 hours of EGF (5 µg/mL) treatment. Staining for EGFR (red), ATP1A1 (green), DAPI (blue), and the overlays of three channels (the membrane localization of EGFR is shown in yellow) are shown in the first, second, third, and fourth rows, respectively (scale bar, 10 µm). (H) Membrane protein fractions of H1975 cotransfected with siRNA-FASN or siRNA-NC and Flag-PKC-iota or empty vector for 48 hours were immunoblotted with anti-p-EGFR-1092 or anti-EGFR. ATP1A1 provided the membrane loading control. (I) H1975 cells were cotransfected with siRNA-FASN or siRNA-NC and Flag-PKC-iota or empty vector for 24 hours. EGF (5 µg/mL) was then added for 24 hours, followed by 30 min of dynasore (80 µM) treatment. Lysates were immunoblotted with anti-FASN, anti-p-EGFR (Tyr 1092), anti-EGFR, or anti-Flag. β-Tubulin provided the loading control. DAPI, 4’,6-diamidino-2-phenylindole; EGF, epidermal growth factor; EGFR, epidermal growth factor receptor; FAs, fatty acids; FASN, fatty acid synthase; GC-MS, gas chromatography-mass spectrometry; IF, immunofluorescence; LFU, lipid fluidity; NC, negative control; PKC-iota, protein kinase C-iota; SFAs, saturated FAs; TG, triglyceride; TMA-DPH, 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluene sulfonate; UFAs, unsaturated FAs.

Article Snippet: The following additional reagents were in the present study: MG132 (HY-13259, MCE, NJ, USA); cycloheximide (HY-12320, MCE); EGFT-TKIs gefitinib (HY-50895, MCE), afatinib (HY-10261, MCE), and osimertinib (HY-15772, MCE); Dynasore (HY-15304, MCE); Pitstop2 (HY-115604, MCE); TMA-DPH (HY-D0986, MCE); EGF (HY-P7109, MCE).

Techniques: Membrane, Plasmid Preparation, Control, Gas Chromatography-Mass Spectrometry, Staining, Transfection, Fluorescence, Gas Chromatography, Mass Spectrometry, Immunofluorescence, Negative Control

Journal: International Journal of General Medicine

Article Title: Characterization of Gastric Mucinous Carcinoma: Clinicopathological Insights and Vascular Features via MSCT

doi: 10.2147/IJGM.S567353

Figure Lengend Snippet: Immunohistochemistry (IHC) detection of vascular endothelial growth factor A (VEGF-A), hypoxia-inducible factor-1α (HIF-1α), and epidermal growth factor (EGF) in carcinomas and corresponding paracancerous normal tissues of MGC, LEMPC, and NMGC. ( A – C ) Representative immunohistochemical staining of VEGF-A ( A ), HIF-1α ( B ), EGF( C ) in carcinomas and normal tissues (×400, bar = 50μm). ( D – F ) IHC score differences (cancer tissue score - normal tissue score) for VEGF-A ( D ) HIF-1α ( E ) EGF ( F ) among the three groups. Statistical significance is indicated as follows: P < 0.05; P < 0.001.

Article Snippet: Sections were stained for CD34 (1:2500, Abcam, ab81289), VEGF-A (1:800, Proteintech, 66828-1-IG), HIF-1α (1:200, Proteintech, 20960-1-AP), and EGF (1:200, Proteintech, 27141-1-AP).

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining

Journal: Indian Journal of Pharmaceutical Sciences

Article Title: Protective Effects of Ginsenoside Rb1 in Rats with Diabetic Cardiomyopathy

doi: 10.36468/pharmaceutical-sciences.967

Figure Lengend Snippet: Fig. 4: Ginsenoside Rb1 attenuated inflammation and suppressed the HB-EGF pathway in DCM rats, (A): RNA expressions of IL6 as determined by quantitative real-time RT-PCR; (B): Protein expressions of HB-EGF in serum as determined by ELISA and (C): Protein expressions of HB-EGF in heart tissue as determined by Western blot Note: Compared to NC, ***p<0.001 and compared to DCM, ###p<0.001

Article Snippet: Human Heparin-Binging Epidermal Growth Factor (HB-EGF) Enzyme-Linked Immunosorbent Assay (ELISA): A Human HB-EGF ELISA kit (Elabscience, China) was used to test for the activity of HB-EGF in plasma, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot