Fig. 1

SNHG5 knockdown inhibits the osteogenic differentiation of hBMSCs. a Relative expression of ALP, RUNX2, OCN, and SNHG5 during osteoinduction of hBMSCs via qRT-PCR (n = 3). GAPDH was used for normalization relative to the day 0 group. b Images of ALP staining on day 7 of osteogenic differentiation, and ARS staining on day 14 of osteogenic differentiation in the si-NC, si-SNHG5-1, and si-SNHG5-2 groups (n = 3). Histograms show ALP activity and ARS staining quantification by spectrophotometry. c Relative mRNA expression of ALP, RUNX2, and OCN measured via qRT-PCR in the PM and OM on day 7. GAPDH was used for normalization (n = 3). d Confocal microscopy of RUNX2 with DAPI counterstaining of the si-NC, si-SNHG5-1, and si-SNHG5-2 groups after osteogenic induction for 7 days (n = 3). Scale bars: 20 μm. e Western blotting analyses of the protein expression of RUNX2, OCN, and β-actin in the si-NC, si-SNHG5-1, and si-SNHG5-2 groups after osteogenic induction for 7 days (n = 3). Histograms show the quantification of band intensities. β-actin was used for normalization relative to the si-NC group. (*p < 0.05; **p < 0.01). ALP, alkaline phosphatase; ARS, alizarin red S; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hBMSCs, human bone marrow mesenchymal stem cells; PM, proliferation medium; OCN, osteocalcin; OM, osteogenic medium; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; RUNX2, runt-related transcription factor 2; SNHG5, small nucleolar RNA host gene 5