aldehyde resin biotin peg3 azide click chemistry  (Chem Impex International)

Journal: The Journal of Clinical Investigation

Article Title: White adipocytes in subcutaneous fat depots require KLF15 for maintenance in preclinical models

doi: 10.1172/JCI172360

Figure Lengend Snippet: ( A ) RT-qPCR quantifying the expression levels of Klf15 in adipocytes isolated from distinct fat pads. 1-way ANOVA, n = 3. ( B ) RT-qPCR quantifying the expression levels of Klf15 in adipocytes after isoproterenol (Iso) treatment for 4 hours. Student’s t test, n = 6. ( C ) RT-qPCR quantifying the expression levels of Klf15 in iWAT isolated from mice after they were injected with CL316243 (CL) (1 mg/kg/day) for 7 days. Student’s t test, n = 4. ( D ) RT-qPCR quantifying the relative expression levels of Adrb1 , Adrb2 , and Adrb3 in iWAT, gWAT, and BAT with iWAT set to 1 for each litter. 1-way ANOVA, n = 5. ( E ) Light phase microscopy images of adipocytes from differentiated SVF harvested from the iWAT of Klf15-fl/fl mice and infected with adenovirus expressing Cre (Ad-Cre) or adenovirus control (Ad-control). Scale bar: 25 μm. ( F ) RT-qPCR quantifying the expression levels of thermogenic genes and panadipocyte marker Adipoq . Student’s t test, n = 4. ( G ) RT-qPCR quantifying the expression levels. Student’s t tests followed by Holm-Šidák correction, n = 7. ( H ) Immunoblots detecting and quantifying the relative levels of β1AR following acute Klf15 deletion in adipocytes. Student’s t test, n = 3. ( I ) Immunoblots detecting and quantifying the relative levels of phosphorylation of p38 MAPK following acute Klf15 deletion in adipocytes. Student’s t test, n = 3. ( J ) RT-qPCR quantifying the change in Ucp1 expression levels with SB202190 pretreatment. Student’s t test, n = 3–4. ( K ) RT-qPCR quantifying the expression levels of Ucp1 following acute Klf15 deletion in adipocytes treated with Isoproterenol. 2-way ANOVA, n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the ADRB1 rescue experiment, adipocytes with hKLF15 knockdown were infected with control shRNA (sc-108080, Santa Cruz Biotechnology) and β1AR shRNA (sc-29580-V, Santa Cruz Biotechnology) lentiviral particles.

Techniques: Quantitative RT-PCR, Expressing, Isolation, Injection, Microscopy, Infection, Control, Marker, Western Blot, Phospho-proteomics

Journal: The Journal of Clinical Investigation

Article Title: White adipocytes in subcutaneous fat depots require KLF15 for maintenance in preclinical models

doi: 10.1172/JCI172360

Figure Lengend Snippet: ( A ) RT-qPCR quantifying the expression levels of Klf15 in iWAT, gWAT, BAT, and liver from WT and Adipo-Klf15 mice. Student’s t tests followed by Holm-Šidák correction, n = 3. ( B ) RT-qPCR quantifying the expression levels of Klf15 in the SVF and adipocyte fraction of iWAT from WT and Adipo-Klf15 mice. Student’s t test followed by Holm-Šidák correction, n = 3. ( C ) Representative images of in situ iWAT in WT and Adipo-Klf15 mice. Scale bar: 5 mm. ( D ) Quantification of iWAT mass as a percent of body weight in WT and Adipo-Klf15 littermates. Ratio paired t test, n = 5. ( E ) RT-qPCR quantifying the expression levels of thermogenic genes in iWAT of WT and Adipo-Klf15 littermates. Student’s t test, n = 5. ( F – H ) RT-qPCR quantifying the expression levels of adrenergic receptors in iWAT, gWAT, and BAT from WT and Adipo-Klf15 mice. Student’s t test followed by Holm-Šidák correction, n = 5. ( I ) RT-qPCR quantifying the expression levels of Adrb1 versus Adrb3 in iWAT in littermates of WT and Adipo-Klf15 mice. 1-tailed ratio paired t test, n = 4. ( J ) Immunoblots detecting the levels of β1AR protein in iWAT from WT and Adipo-Klf15 mice compared with the levels of β-actin controls. ( K ) Quantifying the relative protein level of β1AR. Student’s t test, n = 4. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the ADRB1 rescue experiment, adipocytes with hKLF15 knockdown were infected with control shRNA (sc-108080, Santa Cruz Biotechnology) and β1AR shRNA (sc-29580-V, Santa Cruz Biotechnology) lentiviral particles.

Techniques: Quantitative RT-PCR, Expressing, In Situ, Western Blot

Journal: Scientific Reports

Article Title: Remuscularization with triiodothyronine and β 1 -blocker therapy reverses post-ischemic left ventricular dysfunction and adverse remodeling

doi: 10.1038/s41598-022-12723-2

Figure Lengend Snippet: T3-induced ERK1/2 phosphorylation and cyclin expression in adult ventricular CMs is potentiated by β 1 -AR siRNA. ( A ) Schematic showing the protocol for β 1 -AR siRNA and T3 administration and analysis times. ( B ) Representative immunoblots show expression of indicated proteins in lysates obtained from CMs at 40 h after T3 administration. ( C – M ) Quantitative data obtained from analysis of immunoblots for β 1 -AR ( C ), cyclin D1 ( D ), cyclin A2 ( E ), cyclin B1 ( F ), ECT2 ( G ), p-MEK1/2 ( H ), MEK1/2 ( I ), p-ERK1 ( J ), ERK1 ( K ), p-ERK2 ( L ) and ERK2 ( M ). The data were normalized with loading control (GAPDH) and data were expressed as relative value for scrambled control siRNA without T3 therapy. Immunoblots are representative of 4 biologically independent replicates. Data are mean ± SEM. *** P < 0.001.

Article Snippet: This dose, which was based on dose titration studies , minimizes replication-independent occurrence of monochromatic clusters by ensuring minimal labeling of CMs (~ 1% of the heart) . β 1 -AR-specific siRNA (sc-29581, Santa Cruz Biotechnology) or scrambled siRNA (control) was administered using in vivo-jetPEI, (VWR, 89,129–960). β 1 -AR siRNA (100 ng) was dissolved in 1 ml of the in vivo-jetPEI:10% glucose mixture and was injected 100 μl per mouse via i.p. route (10 ng/mouse). β 1 -AR siRNA was a pool of 2 different siRNA duplexes.

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

Journal: Scientific Reports

Article Title: Remuscularization with triiodothyronine and β 1 -blocker therapy reverses post-ischemic left ventricular dysfunction and adverse remodeling

doi: 10.1038/s41598-022-12723-2

Figure Lengend Snippet: T3-induced ERK1/2 phosphorylation and cyclin expression in adult ventricular CMs is potentiated by β 1 -AR siRNA. ( A ) Schematic showing the protocol for β 1 -AR siRNA and T3 administration and analysis times. ( B ) Representative immunoblots show expression of indicated proteins in lysates obtained from CMs at 40 h after T3 administration. ( C – M ) Quantitative data obtained from analysis of immunoblots for β 1 -AR ( C ), cyclin D1 ( D ), cyclin A2 ( E ), cyclin B1 ( F ), ECT2 ( G ), p-MEK1/2 ( H ), MEK1/2 ( I ), p-ERK1 ( J ), ERK1 ( K ), p-ERK2 ( L ) and ERK2 ( M ). The data were normalized with loading control (GAPDH) and data were expressed as relative value for scrambled control siRNA without T3 therapy. Immunoblots are representative of 4 biologically independent replicates. Data are mean ± SEM. *** P < 0.001.

Article Snippet: This dose, which was based on dose titration studies , minimizes replication-independent occurrence of monochromatic clusters by ensuring minimal labeling of CMs (~ 1% of the heart) . β 1 -AR-specific siRNA (sc-29581, Santa Cruz Biotechnology) or scrambled siRNA (control) was administered using in vivo-jetPEI, (VWR, 89,129–960). β 1 -AR siRNA (100 ng) was dissolved in 1 ml of the in vivo-jetPEI:10% glucose mixture and was injected 100 μl per mouse via i.p. route (10 ng/mouse). β 1 -AR siRNA was a pool of 2 different siRNA duplexes.

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