Journal: bioRxiv

Article Title: MYC-ATF4-ASS1 axis governs intracellular arginine synthesis and dictates the immune microenvironment in melanoma

doi: 10.64898/2026.02.27.707779

Figure Lengend Snippet: ( a ) Western blot to confirm the loss of Ass1 protein expression in YUMM1.7 Ass1 knockout cells (Ass1-KO). β-ACTIN was used as a loading control. ( b ) Ass1-wildtype (Ass1-WT) or Ass1-KO cells were subcutaneously injected into C57BL/6J mice for tumor growth curve. Ass1-KO melanomas does not grow compared to wiltype controls. ( c ) Ass1-WT or Ass1-KO melanomas were collected for histology. IHC staining shows more Cd8+ T cells infiltrated into Ass1-KO melanomas compared to wildtype controls. ( d ) YUMM1.7-WT or Ass1-KO cells were subcutaneously injected into NSG or C57BL/6J respectively for tumor growth curve. Ass1-KO melanomas grow in NSG mice, but not in C57BL/6J mice. ( e ) IHC staining of Cd3 to detect T cells in tumors collected from d. ( f ) YUMM1.7 wiltype cells were subcutaneously injected into C57BL/6J mice and treated with IgG or anti-PD1. Tumors were measured for growth curve. NS indicates no significance.

Article Snippet: Primary antibodies against Ki67 (Thermo Fisher Scientific, MA5-14520), F4/80 (CST, 70076S), Cd8 (98941S), Cd3 (78588S), Cd20 (70168S) were diluted 1:200 in PBST buffer and incubated with slides overnight at 4°C on a shaker.

Techniques: Western Blot, Expressing, Knock-Out, Control, Injection, Immunohistochemistry