preadipocyte growth supplement  (Innoprot Inc)

Journal: Adipocyte

Article Title: Cancer cell-derived exosomal miR-425-3p induces white adipocyte atrophy

doi: 10.1080/21623945.2022.2108558

Figure Lengend Snippet: Exosomal miR-425-3p inhibited preadipocyte proliferation and differentiation. HPA-v cells were incubated with 50 μg/L of exosomes or 50 nM of miR-425-3p mimics for 24 h (for cell proliferation) or 7 days (for adipogenic differentiation), respectively. Adipocyte differentiation was induced using a standard protocol. CytoD (2 μg/mL) was administrated to inhibit endocytosis. Oil red O staining and western blot were performed at day 7 of adipogenic differentiation. (a) Effect of exosomes on cell proliferation. (b) Effect of exosomes on oil red O staining. (c) Effect of exosomes on the protein levels of aP2 and ADPN. (d) Effect of exosomes with miR-425-3p inhibition on cell proliferation. (e) Effect of exosomes with miR-425-3p inhibition on oil red O staining. (f) Effect of exosomes with miR-425-3p inhibition on the protein levels of aP2 and ADPN. (g) Effect of miR-425-3p mimics on cell proliferation. (h) Effect of miR-425-3p mimics on oil red O staining. (i) Effect of miR-425-3p mimics on the protein levels of aP2 and ADPN. Data are presented as mean ± SD, n = 4; ** p < 0.01, *** p < 0.001 vs indicated group. N-Exo: normal (NL20) cell-derived exosomes; C-Exo: cancer (A549) cell-derived exosomes; Mir-Inh-C-Exo: cancer (A549) cell-derived exosomes with miR-425-3p inhibition; CytoD: cytochalasin D; aP2: adipocyte protein 2; ADPN: adiponectin; CNTL: control.

Article Snippet: HPA-v cells were cultured in DMEM containing 5% fetal bovine serum (FBS), 1% preadipocyte growth supplement (#7252, ScienCell Research Laboratories), and 1% penicillin/streptomycin (P/S).

Techniques: Incubation, Staining, Western Blot, Inhibition, Derivative Assay, Control

Journal: Adipocyte

Article Title: Cancer cell-derived exosomal miR-425-3p induces white adipocyte atrophy

doi: 10.1080/21623945.2022.2108558

Figure Lengend Snippet: PDE4B depletion did not affect preadipocyte proliferation and differentiation, but promoted adipocyte lipolysis and white adipocyte browning. HPA-v cells were infected with lentiviruses carrying PDE4B shRAN to reduce PDE4B expression. Adipocyte differentiation was induced using a standard protocol. Oil red O staining and western blot were performed at day 7 of adipogenic differentiation. (a) Cell proliferation. (b) The protein levels of GATA2, IGFBP4, and MMP4 in PDE4B KD or control HPA-v cells. (c) Oil red O staining. (d) The protein levels of aP2 and ADPN in differentiated cells. To observe the potential effects of PDE4B on adipocyte lipolysis and white adipocyte browning, lentiviruses carrying PDE4B shRAN were used to knockdown PDE4B in mature adipocyte. (e) Oil red O staining. (f) Glycerol concentration in culture medium. (g) UCP1 protein levels. Data are presented as mean ± SD, n = 4; ** p < 0.01, *** p < 0.001 vs indicated group. KD: knockdown; aP2: adipocyte protein 2; ADPN: adiponectin; CNTL: control, ns: no significant.

Article Snippet: HPA-v cells were cultured in DMEM containing 5% fetal bovine serum (FBS), 1% preadipocyte growth supplement (#7252, ScienCell Research Laboratories), and 1% penicillin/streptomycin (P/S).

Techniques: Infection, Expressing, Staining, Western Blot, Control, Knockdown, Concentration Assay

Journal: Adipocyte

Article Title: Cancer cell-derived exosomal miR-425-3p induces white adipocyte atrophy

doi: 10.1080/21623945.2022.2108558

Figure Lengend Snippet: A schematic diagram for the impacts of cancer cell-derived exosomal miR-425-3p on adipocyte atrophy. Cancer cell-derived exosomal miR-425-3p targets proliferation- and differentiation-related regulating genes leading to the inhibition of preadipocyte proliferation and differentiation. In addition, PDE4B downregulation induced by cancer cell-derived exosomal miR-425-3p results in enhancement of adipocyte lipolysis and white adipocyte browning through activating cAMP/PKA signalling and/or lipophagy, respectively. The combined effects of these alterations will bring about adipocyte atrophy, ultimately leading to the loss of adipose tissue in cancer cachexia.

Article Snippet: HPA-v cells were cultured in DMEM containing 5% fetal bovine serum (FBS), 1% preadipocyte growth supplement (#7252, ScienCell Research Laboratories), and 1% penicillin/streptomycin (P/S).

Techniques: Derivative Assay, Inhibition