Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: Expression profile of TRPC6 in cold stimulation and mature adipocytes of BAT. ( A ) Wildtype mice were transferred to 4 °C for up to 7 d. ( B ) Representative H&E staining of BAT at room temperature (RT) and cold exposure (Scale bar, 100 μm). ( C ) Lipid droplet area (n = 8). ( D ) Volcano plot of differential genes by RNA-seq. ( E ) GO biological process analysis. ( F ) Relative expression of TRPC family genes differentially expressed (RT and 4 °C, n = 3). ( G ) The mRNA levels of Trpc6 in BAT after 7 days cold exposure (n = 3). Statistical comparisons were made between D1 vs. D0, D4 vs. D0, and D7 vs. D0 using one-way ANOVA. ( H ) The protein levels and (I ) quantification of protein levels of TRPC6, UCP1 and PGC-1α in BAT (n = 5). ( J ) IHC staining (TRPC6) of BAT (Scale bar, 100 μm). ( K ) Quantitative analysis of IHC staining (TRPC6) of BAT (n = 7). ( L ) Fluorescence staining of primary brown adipocytes (Scale bar, 50 μm). ( M ) The mRNA level of Trpc6 in adipocytes and SVFs of BAT (n = 3). ( N ) Fluorescence staining of C3H10T1/2-derived adipocytes (Scale bar, 100 μm). ( O ) The protein levels and ( P ) quantification of protein levels of TRPC6, UCP1 and PGC-1α in preadipocytes (D0, D1, and D3) and adipocytes (D1, D3, D5, and D7) (n = 3). ( Q ) Trpc6 mRNA level in adipocytes and preadipocytes of C3H10T1/2 cells (n = 3). Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant by unpaired two-tailed Student's t tests.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: Expressing, Staining, RNA Sequencing, Immunohistochemistry, Fluorescence, Derivative Assay, Two Tailed Test

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: TRPC6 deficiency in BAT aggravates HFD-induced Obesity. ( A – H ) Trpc6 fl/fl and Trpc6 BTKO mice were fed normal chow diet (NCD) for 9 weeks. ( A ) Representative images of mice. ( B ) Body weight (n = 6). ( C ) Averaged weekly food intake (n = 9 weeks/cage). ( D ) Representative images of BAT. ( E ) Relative BAT weight (n = 6). ( F ) Representative H&E stains of BAT (Scale bar, 100 μm). ( G ) Lipid droplet area distribution and ( H ) means in BAT (n = 7–8). ( I – V ) Trpc6 fl/fl and Trpc6 BTKO mice were fed high-fat diet (HFD) for 14 weeks. ( I ) Representative appearance of mice. ( J ) Body weight (n = 6). ( K ) Body composition of mice (n = 6). ( L ) Representative images of BAT. ( M ) Relative weights of adipose tissue and liver (n = 7–10). ( N ) Representative H&E stains of BAT (Scale bar, 100 μm). ( O ) Lipid droplet area distribution and ( P ) means in BAT (n = 7–8). ( Q ) Representative H&E stains of liver (Scale bar, 200 μm). ( R , T ) Glucose tolerance test (GTT) and insulin tolerance test (ITT) (n = 6). ( S , U ) Analysis of the GTT and ITT data using an area of the curve (n = 6). Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques:

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: TRPC6 deficiency impairs energy balance and cold tolerance. ( A ) Averaged weekly food intake of Trpc6 fl/fl and Trpc6 BTKO mice fed HFD (n = 13 weeks/cage). ( B ) Energy expenditure (n = 4). ( C ) Oxygen consumption (n = 4). ( D ) Carbon dioxide production (n = 4). ( E ) Respiratory exchange ratio (n = 4). ( F ) Trpc6 fl/fl and Trpc6 BTKO mice were transferred to 4 °C for up to 7 d. ( G ) Core body temperatures upon acute 4 °C exposure (n = 6). ( H ) Representative forward-looking infra-red (FLIR) image and ( I ) analysis of capturing surface body temperature at 2 h acute cold exposure (n = 6). ( J ) Representative images of BAT after 7 days cold exposure. ( K ) Relative BAT weight (n = 6). ( L ) Representative H&E stains of BAT (Scale bar, 100 μm). ( M ) Lipid droplet area distribution and ( N ) means in BAT (n = 6–7). Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant by unpaired two-tailed Student's t tests.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: Two Tailed Test

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: TRPC6 deficiency suppresses the thermogenic program via PGC-1α/UCP1 signaling in BAT. ( A – G ) Trpc6 fl/fl and Trpc6 BTKO mice were transferred to 4 °C for up to 7 d. ( A ) The protein levels and ( B ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin, and OXPHOS complex proteins in BAT (n = 6). ( C ) The mRNA levels of thermogenic genes in BAT (n = 6). ( D ) IHC staining (UCP1 and PGC-1α) of BAT (Scale bar, 100 μm). ( E ) Quantitative analysis of IHC staining (UCP1 and PGC-1α) of BAT (n = 7). ( F ) Relative mitochondrial DNA (mtDNA) levels (n = 8). ( G ) Representative TEM images of mitochondria in BAT (Top, Scale bar, 1 μm; Bottom, Scale bar, 1 μm). ( H – N ) Trpc6 fl/fl and Trpc6 BTKO mice were fed HFD for 14 weeks. ( H ) The protein levels and ( I ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin, and OXPHOS complex proteins in BAT (n = 6). ( J ) The mRNA levels of thermogenic genes in BAT (n = 7–9). ( K ) IHC staining (UCP1 and PGC-1α) of BAT (Scale bar, 100 μm). ( L ) Quantitative analysis of IHC staining (UCP1 and PGC-1α) of BAT (n = 8). ( M ) Relative mtDNA levels (n = 8). ( N ) Representative TEM images of mitochondria in BAT (Top, Scale bar, 1 μm; Bottom, Scale bar, 1 μm). Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant by unpaired two-tailed Student's t tests.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: Immunohistochemistry, Two Tailed Test

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: Impact of TRPC6 on adipocytes in vitro. ( A ) Representative fluorescence staining of primary brown adipocytes from Trpc6 +/+ and Trpc6 −/− (Scale bar, 50 μm). ( B – C ) Lipid droplets area (n = 11). ( D ) Ca 2+ influx was detected by using the Ca 2+ indicator Fluo-4 AM and ( E ) Ca 2+ relative fluorescence intensity (n = 6). ( F ) The protein levels and ( G ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin, and OXPHOS complex proteins (n = 3). ( H ) The mRNA levels of thermogenic genes (n = 4). ( I ) Oxygen consumption rate of Mito-stress test and ( J ) assay parameters of Mito-stress test (n = 9). ( K ) Representative Oil Red O-stained (top; Scale bar, 200 μm) and phase contrast images (bottom; Scale bar, 100 μm), and ( L ) quantitative analysis of lipid droplet area of C3H10T1/2-derived adipocytes infected with sh- Trpc6 or vector (Ctrl) (n = 10–18). ( M ) The protein levels and ( N ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin, and OXPHOS complex proteins (n = 3). ( O ) The mRNA levels of thermogenic genes (n = 3). ( P ) Representative fluorescence staining (Scale bar, 100 μm) and ( Q ) lipid droplets area analysis of C3H10T1/2-derived adipocytes transduced with Trpc6 OE or vector (Ctrl) (n = 15). ( R ) The protein levels and ( S ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin and OXPHOS complex proteins (n = 3). ( T ) The mRNA levels of thermogenic genes (n = 3). ( U ) Representative fluorescence images of mitochondrial morphology (Scale bar, 10 μm). ( V ) Quantitative analysis of mitochondrial density (n = 5). Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant by unpaired two-tailed Student's t tests.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: In Vitro, Fluorescence, Staining, Derivative Assay, Infection, Plasmid Preparation, Transduction, Two Tailed Test

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: Knockout of TRPC6 in BAT impairs thermogenesis by inhibiting the p38-MAPK pathway. ( A ) KEGG enrichment process analysis of RNA-seq of Trpc6 fl/fl and Trpc6 BTKO mice after cold exposure (n = 3). ( B ) The protein levels and ( C ) quantification of protein levels of TRPC6, β-Actin, and p38 MAPK in BAT after cold exposure (n = 6). ( D ) The protein levels and ( E ) quantification of protein levels of TRPC6, β-Actin, and p38 MAPK in BAT after HFD (n = 6). ( F ) The protein levels and ( G ) quantification of protein levels of TRPC6, β-Actin, and MAPK in primary brown adipocytes (n = 3). ( H ) The protein levels and ( I ) quantification of protein levels of TRPC6, β-Actin, and MAPK in C3H10T1/2-derived adipocytes infected with sh- Trpc6 or vector (n = 3). ( J ) The protein levels and ( K ) quantification of protein levels of TRPC6, β-Actin, and MAPK in C3H10T1/2-derived adipocytes infected with Trpc6 OE or vector (n = 3). ( L – M ) Primary brown adipocytes loaded with Fluo-4 AM. Baseline [Ca 2+ ] i was acquired. Primary brown adipocytes, incubated in Ca 2+ -containing or Ca 2+ -free bath solutions, were exposed to HPF (5 μM) at 150 s. ( L ) A representative experiment was described where fluorescence was represented as F/F 0 as a function of time. ( M ) The first peak (n = 3). ( N – O ) C3H10T1/2-derived adipocytes loaded with Fluo-4 AM. ( N ) A representative experiment was described where fluorescence was represented as F/F 0 as a function of time. ( O ) The first peak (n = 3). ( P – Q ) Primary brown adipocytes were treated with vehicle (Ctrl) or HPF for 48 h. ( P ) The protein levels and ( Q ) quantification of protein levels of UCP1, PGC-1α, β-Actin, and p38 MAPK (n = 3). ( R – S ) Trpc6 −/− adipocytes were treated with vehicle or HPF for 48 h. ( R ) The protein levels and ( S ) quantification of protein levels of UCP1, PGC-1α, β-Actin, and p38 MAPK (n = 3). Unpaired two-tailed Student's t tests and one-way ANOVA were used appropriately. Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: Knock-Out, RNA Sequencing, Derivative Assay, Infection, Plasmid Preparation, Incubation, Fluorescence, Two Tailed Test

Journal: Molecular Metabolism

Article Title: TRPC6 governs brown adipose thermogenesis via a BMPR2-p38 MAPK signaling axis

doi: 10.1016/j.molmet.2026.102341

Figure Lengend Snippet: TRPC6 knockdown suppresses thermogenic program through BMPR2. ( A ) Protein–protein interaction (PPI) network of TRPC6 predicted by STRING database. ( B – C ) Surface electrostatic potential of TRPC6 and BMPR2 (Blue: positive charge, Red: negative charge). ( D ) Molecular docking analysis indicated possible binding interactions between TRPC6 and BMPR2 proteins (Purple: TRPC6, Turquoise: BMPR2). ( E ) Immunoprecipitation confirmed the interaction between TRPC6 and BMPR2. ( F ) Representative fluorescence staining and ( G ) gray value of TRPC6 and BMPR2 in BAT (Red: TRPC6, Green: BMPR2, Blue: DAPI; Scale bar, 10 μm). ( H ) The protein levels and ( I ) quantification of protein levels of TRPC6, BMPR2, and β-Actin in primary brown adipocytes from Trpc6 +/+ and Trpc6 −/− (n = 3). ( J ) The protein levels and ( K ) quantification of protein levels of TRPC6, BMPR2, and β-Actin in C3H10T1/2-derived adipocytes infected with sh- Trpc6 or vector (n = 3). ( L ) The protein levels and ( M ) quantification of protein levels of TRPC6, BMPR2, and β-Actin in C3H10T1/2-derived adipocytes infected with Trpc6 OE or vector (n = 3). ( N ) Representative fluorescence staining (Scale bar, 100 μm) and ( O ) lipid droplets area analysis of C3H10T1/2-derived adipocytes transfected with Bmpr2 siRNA (si- Bmpr2 ) or vector (Ctrl) (n = 11–12). ( P ) The protein levels and ( Q ) quantification of protein levels of TRPC6, UCP1, PGC-1α, β-Actin, and OXPHOS complex proteins (n = 3). ( R ) The protein levels and ( S ) quantification of protein levels of BMPR2, β-Actin, and p38 MAPK (n = 3). ( T – U ) C3H10T1/2-derived adipocytes of Trpc6 OE transfected with Bmpr2 siRNA or vector. ( T ) The protein levels and ( U ) quantification of protein levels of TRPC6, BMPR2, UCP1, PGC-1α, β-Actin, and p38 MAPK (n = 3). ( V ) The protein levels and ( W ) quantification of protein levels of UCP1, PGC-1α, β-Actin, and p38 MAPK in primary brown adipocytes following 48 h treatment with vehicle (Ctrl) or DHC (1 μM) (n = 3). ( X–Y ) Trpc6 −/− adipocytes were treated with vehicle or DHC for 48 h. ( X ) The protein levels and ( Y ) quantification of protein levels of UCP1, PGC-1α, β-Actin, and p38 MAPK (n = 3). Unpaired two-tailed Student's t tests and one-way ANOVA were used appropriately. Data are presented as mean ± SEM and ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, ns, not significant.

Article Snippet: To knockdown Trpc6 , sh -Trpc6 was generated by GeneChem Co., Ltd (Shanghai, China).

Techniques: Knockdown, Binding Assay, Immunoprecipitation, Fluorescence, Staining, Derivative Assay, Infection, Plasmid Preparation, Transfection, Two Tailed Test