Journal: Nature Communications

Article Title: PRMT3-mediated post-translational adaptation to fasting regulates metabolic flexibility

doi: 10.1038/s41467-026-68883-6

Figure Lengend Snippet: a , b Western blot analysis ( a ) and quantification ( b ) of SLC25A1 at d0, d2, d4 and d8 during adipogenic differentiation of preadipocytes isolated from iWAT ( n = 3). c Relative levels of SLC25A1 in eWAT of 20 weeks old male mice fed with 12 weeks of normal diet (ND) or high-fat diet (HFD) ( n = 4). d Western blot results show the specific deletion of SLC25A1 in eWAT but not liver from Slc25a1 AKO mice, representative image from 2 independent repeats. e Body weight of male WT ( Slc25a1 -flox/flox) and Slc25a1 AKO (Adiponectin-Cre; Slc25a1 flox/flox) mice during 10 weeks with HFD ( n = 10 and 11). f Body composition of WT and Slc25a1 AKO mice after 10 weeks of HFD ( n = 5 and 7). g , h Blood glucose levels and area under the curve (AUC) or area above the curve (AAC) of male WT and Slc25a1 AKO mice during glucose tolerance test or insulin tolerance test after 10 weeks of HFD, respectively ( n = 10 and 11). i Tissue weights of various adipose tissues, liver and tibial anterior muscle (TA) after 10 weeks of HFD ( n = 6 and 8). j Representative H&E staining image of liver and eWAT from WT and Slc25a1 AKO mice after HFD from 4 pairs of animals, Bar = 250 μm. k Levels of cholesterol, HDL, LDL, and TG from the serum of WT and Slc25a1 AKO mice after HFD ( n = 5). l Blood glucose of WT and Slc25a1 AKO mice after HFD, fasted from zt1 to zt7 ( n = 10 and 11). m Seahorse analysis of ECAR in mouse eWAT-derived adipocytes treated with or without CTPI-2 ( n = 5). n Quantification of the glycolic ability of cultured eWAT adipocyte with or without CTPI-2 treatment from ( m ). o Levels of compensatory glycolysis from cultured adipocytes from Ctrl, CTPI-2 treated, and Slc25a1 AKO mice ( n = 6, 6 and 7). p Gene Ontology (GO) annotation to identify the key up-regulated pathways in eWAT of Slc25a1 AKO mice compared to WT. q Heatmap of DEGs enriched in glucose metabolism. r , s Western blot analysis ( r ) and quantification ( s ) of SLC25A1, GAPDH, PC and PKM1/2 in eWAT of WT and Slc25a1 AKO mice ( n = 3). t 48 h VO 2 rhythms and average VO 2 levels at indicated time frames ( n = 7 and 10). Data are mean ± s.e.m. Statistical significance was determined using unpaired two-tailed Student’s t test. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used for immunoblots were as follows: anti-Mono-Methyl Arginine (8015, MMA, diluted 1:1000), Asymmetric Di-Methyl Arginine Motif (13522, ADMA, diluted 1:1000), Symmetric Di-Methyl Arginine Motif (13222, SDMA, diluted 1:1000), Normal anti-rabbit lgG (2729), Normal anti-mouse lgG (68860), Phospho-Akt (Ser473) (4060, diluted 1:1000), AKT (9272, diluted 1:1000), PRMT4 (3379, diluted 1:1000), PRMT5 (79998, diluted 1:1000), PKM1/2 (3106, diluted 1:1000), Pyruvate Carboxylase (49381, PC, diluted 1:1,000), LDHA (2012, diluted 1:1000), CPT2 (52552, diluted 1:1000), PGK1 (63536, diluted 1:1000), PCK1 (12940, diluted 1:1000), Phospho-HSL (Ser660) (45804, diluted 1:1000) antibodies were purchased from Cell Signaling technology.

Techniques: Western Blot, Isolation, Staining, Derivative Assay, Cell Culture, Two Tailed Test

Journal: Journal of Clinical Investigation

Article Title: ST6GalNAc-I regulates tumor cell sialylation via NECTIN2/MUC5AC-mediated immunosuppression and angiogenesis in non–small cell lung cancer

doi: 10.1172/jci186863

Figure Lengend Snippet: Figure 2. Overexpression of NECTIN2 in LUAD and tumor cell sialylation. (A and B) In silico analysis indicated that expression of NECTIN2 is significantly overexpressed and associated with poor survival outcomes in both early- and late-stage LUAD patients. (C) Protein-protein interaction networking analysis reveals that tumor cells expressing NECTIN2 induce T cell dysfunction through TIGIT binding, which is associated with immune suppression pathways. (D) IHC analysis shows the overexpression of NECTIN2 in LUAD. Data were analyzed using 2-tailed t test (n = 38). Original magnification, ×10. (E) The expression of MUC5AC and NECTIN2 drastically decreased in ST6GalNAc-I–KO and MUC5AC-KD cells (A549 and H1437). (F and G) Immunoprecipitation assay shows NECTIN2 sialylation in LUAD cells. (H) SNA pull-down was performed on A549 cell lysates, followed by immunoblotting with NECTIN2 antibody, suggesting that NECTIN2 carries STn in LUAD cells. (I) Immunofluorescence assay reveals the decreased association of NECTIN2 and STn in A549 ST6GalNAc-I–KO cells, and the bar diagram represents the quantification of NECTIN2 and STn using arith- metic mean intensity. Data were analyzed using 2-tailed t test (n = 3). Scale bars: 5 μm.

Article Snippet: Immunoprecipitation was carried out using NECTIN2 (catalog 27171-1-AP, ProteinTech) per the abovementioned protocol and detected using TIGIT (catalog NBP2-79794, Novus Biologicals).

Techniques: Over Expression, In Silico, Expressing, Binding Assay, Immunoprecipitation, Western Blot, Immunofluorescence

Journal: Journal of Clinical Investigation

Article Title: ST6GalNAc-I regulates tumor cell sialylation via NECTIN2/MUC5AC-mediated immunosuppression and angiogenesis in non–small cell lung cancer

doi: 10.1172/jci186863

Figure Lengend Snippet: Figure 4. St6galnac-I–associated immunosuppression in LUAD. (A) We developed stable mouse St6galnac-I knockdown in mouse syngeneic KP2075 cells. Quantitative real-time PCR analysis shows that the transcript level of St6galnac-I was significantly decreased in St6galnac-I–KD cells. Significance was determined by 2-tailed t test (n = 4). (B) Immunoblot shows that Nectin2 and Muc5ac were decreased upon St6galnac-I knockdown. (C) Schemes for the intratracheal orthotopic models. (D–F) Mice injected with St6galnac-I–KD cells showed reduced lung tumor incidence in H&E (n = 6) along with decreased Ki-67, St6galnac-I, STn, Nectin2, and Tigit. Significance was determined by 2-tailed t test (n = 3). Original magnification, ×4 for H&E and ×40 for IHC. (G) Left: Immunofluorescence assays indicate decreased association of Nectin2 and Tigit in St6galnac-I–KD tumors. Right: Quantification of the arithmetic mean intensity value of CD3 (green), NECTIN2 (red), and TIGIT (purple) per field of view (n = 3). Scale bars: 5 μm.

Article Snippet: Immunoprecipitation was carried out using NECTIN2 (catalog 27171-1-AP, ProteinTech) per the abovementioned protocol and detected using TIGIT (catalog NBP2-79794, Novus Biologicals).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Injection, Immunofluorescence