cebpa cell signaling 2295s antibody  (Cell Signaling Technology Inc)

Journal: Scientific reports

Article Title: miR-181a modulates circadian rhythm in immortalized bone marrow and adipose derived stromal cells and promotes differentiation through the regulation of PER3.

doi: 10.1038/s41598-018-36425-w

Figure Lengend Snippet: Figure 1. miR-181a is induced during BMSC/ASC adipogenesis. (A) Graphs depicting increasing expression of miR-181a during the induction of adipogenesis at days 0, 1, 3, and 7 for Scp-1 and PASC-1 cells. (B) Graphs depicting increasing mRNA expression of PPARG, CEBPA, and terminal adipogenesis markers during the induction of adipogenesis at days 0, 1, 3, and 7 for Scp-1 and PASC-1 cells. (C) Representative western blot images showing increasing protein expression of PPARG, CEBPA, and FABP4 during the induction of adipogenesis at days 0, 1, 3, and 7 for Scp-1 cells with respective quantifications below (Full blots are shown in Supplemental Fig. 5A). (D) Representative western blot images showing increasing protein expression of PPARG, CEBPA, and FABP4 during the induction of adipogenesis at days 0, 1, 3, and 7 for PASC-1 cells with respective quantifications below (Full blots are shown in Supplemental Fig. 5B). (E) Micrographs showing Oil red O staining at days 0 and 14 for Scp-1 and PASC-1 cells. Data are representative of 3 independent experiments. * is p ≤ 0.05, ** is p ≤ 0.005, *** is p ≤ 0.0005.

Article Snippet: Primary Antibody Company Catalog # Dilution PPARG Cell Signaling Technology 2443 1:500 CEBPA Cell Signaling Technology 2295 1:500 FABP4 Cell Signaling Technology 3544 1:500 PER3 Abcam ab201940 1:500 GAPDH Santa Cruz sc-365062 1:2500 Secondary Antibody Company Catalog # Mouse anti-rabbit Cell Signaling Technology 58802 1:2500 Rabbit anti mouse Cell Signaling Technology 7076 1:2500 Table 2.

Techniques: Expressing, Western Blot, Staining

Journal: Scientific reports

Article Title: miR-181a modulates circadian rhythm in immortalized bone marrow and adipose derived stromal cells and promotes differentiation through the regulation of PER3.

doi: 10.1038/s41598-018-36425-w

Figure Lengend Snippet: Figure 2. Overexpression of miR-181a in Scp-1 cells increases adipogenesis. (A) Oil red O staining for Scp-1 cells overexpressing either scramble vector (p000) or miR-181a (p181a) on day 14 of adipogenic differentiation with representative micrographs (top) and quantification of Oil red O staining for Scp-1 p000 and p181a cells at day 14 (bottom). (B) Graph depicting increased expression of miR-181a at days 0 and 7 in Scp-1 p181a vs p000 cells. (C) Graphs depicting increased mRNA expression of PPARG and CEBPA at day 0 and increased expression of adipogenesis target genes at day 7 in Scp-1 p181a vs p000 cells. (D) Representative western blot images showing increased expression of CEBPA and PPARG at day 0 in Scp-1 p181a vs p000 cells with quantification (right) (Full blots are shown in Supplemental Fig. 6A). (E) Representative western blot images showing increased expression of CEBPA, PPARG, and FABP4 at day 7 in Scp-1 p181a vs p000 cells with quantification (right) (Full blots are shown in Supplemental Fig. 6B). Data are representative of 3 independent experiments. * is p ≤ 0.05, ** is p ≤ 0.005, *** is p ≤ 0.0005.

Article Snippet: Primary Antibody Company Catalog # Dilution PPARG Cell Signaling Technology 2443 1:500 CEBPA Cell Signaling Technology 2295 1:500 FABP4 Cell Signaling Technology 3544 1:500 PER3 Abcam ab201940 1:500 GAPDH Santa Cruz sc-365062 1:2500 Secondary Antibody Company Catalog # Mouse anti-rabbit Cell Signaling Technology 58802 1:2500 Rabbit anti mouse Cell Signaling Technology 7076 1:2500 Table 2.

Techniques: Over Expression, Staining, Plasmid Preparation, Expressing, Western Blot

Journal: Scientific reports

Article Title: miR-181a modulates circadian rhythm in immortalized bone marrow and adipose derived stromal cells and promotes differentiation through the regulation of PER3.

doi: 10.1038/s41598-018-36425-w

Figure Lengend Snippet: Figure 4. Knockdown of PER3 phenocopies miR-181a overexpression in Scp-1 MSCs. (A) Graph showing miR-181a levels at days 0 and 7 of adipogenic differentiation for Scp-1 cells overexpressing either scramble vector (p000) or miR-181a (p181a) or Scp-1 cells expressing PER3 shRNAs (shPER3 #1 & shPER3 #2). (B) Graph showing PER3 mRNA levels at days 0 and 7 of adipogenic differentiation in Scp-1 p000, p181a, shPER3 #1, and shPER3 #2 cells. (C) Western blot showing PER3 protein levels at days 0 and 7 of adipogenic differentiation in Scp-1 p000, p181a, shPER3 #1, and shPER3 #2 cells (Full blots are shown in Supplemental Fig. 8A,B). (D) Oil red O staining images of Scp-1 p000, p181a, shPER3 #1, and shPER3 #2 cells on day 14 of adipogenic differentiation with representative micrographs (top) and quantification of Oil red O staining (bottom). (E) Graphs depicting increased mRNA expression of PPARG and CEBPA at day 0 and increased expression of adipogenesis target genes at day 7 in Scp-1 p181a, shPER3 #1, and shPER3 #2 vs p000 cells. (F) Representative western blot images showing increased expression of CEBPA and PPARG in Scp-1

Article Snippet: Primary Antibody Company Catalog # Dilution PPARG Cell Signaling Technology 2443 1:500 CEBPA Cell Signaling Technology 2295 1:500 FABP4 Cell Signaling Technology 3544 1:500 PER3 Abcam ab201940 1:500 GAPDH Santa Cruz sc-365062 1:2500 Secondary Antibody Company Catalog # Mouse anti-rabbit Cell Signaling Technology 58802 1:2500 Rabbit anti mouse Cell Signaling Technology 7076 1:2500 Table 2.

Techniques: Knockdown, Over Expression, Plasmid Preparation, Expressing, Western Blot, Staining

Journal: Scientific reports

Article Title: miR-181a modulates circadian rhythm in immortalized bone marrow and adipose derived stromal cells and promotes differentiation through the regulation of PER3.

doi: 10.1038/s41598-018-36425-w

Figure Lengend Snippet: Figure 5. Knockdown of endogenous miR-181a inhibits adipogenesis in BM-MSCs. (A) Graph showing miR- 181a levels at days 0 and 21 of adipogenic differentiation for Scp-1 cells overexpressing either control vector (cmiR) or miR-181a specific antagomiR (antimiR). (B) Oil red O staining images of Scp-1 cmiR and antimiR cells on day 21 of adipogenic differentiation with representative micrographs (top) and quantification of Oil red O staining (bottom). (C) Graphs depicting decreased mRNA expression of PPARG and CEBPA at day 0 and decreased expression of adipogenesis target genes at day 21 in Scp-1 antimiR vs cmiR cells. (D) Representative western blot images showing decreased expression of CEBPA and PPARG in Scp-1 antimiR vs cmiR cells at day 0 with quantification (right) (Full blots are shown in Supplemental Fig. 9A). (E) Representative western blot images showing decreased expression of CEBPA, PPARG, and FABP4 at day 21 in Scp-1 antimiR vs cmiR cells with quantification (right) (Full blots are shown in Supplemental Fig. 9B). (F) Graph showing PER3 mRNA levels at days 0 and 21 of adipogenic differentiation in Scp-1 cmiR and antimiR cells. (G) Western blot showing PER3 protein levels at days 0 and 21 of adipogenic differentiation in Scp-1 cmiR and antimiR cells (Full blots are shown in Supplemental Fig. 9C,D). Data are representative of 3 independent experiments. * is p ≤ 0.05, ** is p ≤ 0.005, *** is p ≤ 0.0005.

Article Snippet: Primary Antibody Company Catalog # Dilution PPARG Cell Signaling Technology 2443 1:500 CEBPA Cell Signaling Technology 2295 1:500 FABP4 Cell Signaling Technology 3544 1:500 PER3 Abcam ab201940 1:500 GAPDH Santa Cruz sc-365062 1:2500 Secondary Antibody Company Catalog # Mouse anti-rabbit Cell Signaling Technology 58802 1:2500 Rabbit anti mouse Cell Signaling Technology 7076 1:2500 Table 2.

Techniques: Knockdown, Control, Plasmid Preparation, Staining, Expressing, Western Blot