Journal: bioRxiv

Article Title: miR-378a and NPNT coordinate autophagy regulation in podocytes through mTOR and MAPK signaling

doi: 10.64898/2026.03.19.709781

Figure Lengend Snippet: a-c: Western blot analysis of EGFR (a), AKT (b) and ERK1/2 (c) phosphorylation in podocytes following NPNT siRNA or control siRNA transfection. Phospho-protein levels were normalized to total protein levels, respectively. Vinculin (a) or GAPDH (b, c) served as loading controls. Data is presented as fold change relative to control siRNA (Ctrl). Data represent mean ± SD, n = 3-5 independent experiments. * p <0.05.

Article Snippet: Primary antibodies used in this study: Phospho-Akt (Ser473, #9271S, Cell Signaling), AKT (#9272, Cell Singaling), phospho-EGFR (Tyr1068, #3777S, Cell Signaling), EGFR (#2232, Cell Signaling), phospho-ERK1/2 (phosphor-p44/42 MAPK (Thr202/Tyr204), #9106, Cell Signaling), ERK1/2 (p44/42 MAPK, #9102, Cell Signaling), GAPDH (sc-32233, Santa Cruz), LC3 (NB100-2220, Novus), phospho-mTOR (Ser2448, #5536S, Cell Signaling), mTOR (2983S, Cell Signaling), vinculin (V9131, Sigma-Aldrich).

Techniques: Western Blot, Phospho-proteomics, Control, Transfection

Journal: ImmunoTargets and Therapy

Article Title: Kasugamycin Inhibits Melanoma Lung Metastasis and CHI3L1-Driven M2-Like Tumor-Associated Macrophage Differentiation

doi: 10.2147/ITT.S563951

Figure Lengend Snippet: KSM inhibits CHI3L1-stimulated M2 macrophage activation via EGFR. Bulk RNA-seq sequencing analysis on the differentiated macrophages from THP-1 cells was used to evaluate mRNA expression in the macrophage differentiation regulated by CHI3L1 and KSM. ( A ). Volcano plots showing differentially expressed genes regulated by CHI3L1. ( B ) Representative plots of the top 20 genes that are upregulated (> 2-fold) by CHI3L1 stimulation but downregulated (< 2-fold) by KSM treatment. ( C ) Representative mRNA expression of EGFR in differentiated macrophages with CHI3L1 and KSM treatment was detected by real-time qRT-PCR. ( D ) Representative immunoblots showing p-EGFR (Tyr1068) and total EGFR expression in differentiated macrophages with CHI3L1 and KSM. Right panel, densitometric quantitation on the blots of EGFR and p-EGFR. ( E ) Effect of EGFR inhibition on the expression of p-STAT3 and total-STAT3 in CHI3L1-stimulated, differentiated macrophages by gefitinib (Tocris Bioscience, #3000) treatment (1 μM, 72 hours). Right panel, densitometric quantitation on the blots of p-STAT3 and total STAT3. ( F ) Effects of EGFR inhibition on the expression of CD163 and CD206 in CHI3L1-stimulated, differentiated macrophages by gefitinib treatment (0.1 and 1 μM, 72 hours). Right panel, densitometric quantitation on the blots of CD163 and CD206. The values in ( C – F ) are the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001 (One-Way ANOVA, multiple comparisons).

Article Snippet: Protein lysates (25 μg) from lung tissues or cells were subjected to SDS-PAGE, transferred to membranes, and immunoblotted with primary antibodies against CD163 (#PA5-109327, Thermo Fisher Scientific), CD206 (E6T5J, #24595S, Cell Signaling Technology), phosphorylated AKT (p-Akt) (193H12, #4058S, Cell Signaling Technology), and total Akt (11E7, #4685S, Cell Signaling Technology), phosphorylated EGFR (Tyr1068) (p-EGFR) (#44-788G, Thermo Fisher Scientific), EGFR (D38B1, #4267S, Cell Signaling Technology), phosphorylated Erk (p-Erk) (#9101S, Cell Signaling Technology), total Erk (#9102S, Cell Signaling Technology), phosphorylated STAT3 (Tyr705) (p-STAT3) (D3A7, #9145, Cell Signaling Technology), total STAT3 (79D7, #4904, Cell Signaling Technology) and b-actin (C4, #sc-47778 HRP, Santa Cruz Biotechnology).

Techniques: Activation Assay, RNA Sequencing, Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Quantitation Assay, Inhibition