SNHG12 Promotes Autophagy by Blocking the mTOR-Primary Cilia-mTOR Loop via Activating the miR-181a-5p/miR-138-5p-INPP5E Axis in Chondrocyte

Figure 4

Silencing SNHG12 promotes primary cilia and autophagy via the hsa-miR-181a-5p/hsa-miR-138-5p-INPP5E axis in vitro. (a, b) RT-PCR and WB assay were used to detect the expression of INPP5E in control, NC, and OE-INPP5E groups. (c, d) RT-PCR and WB assay were used to detect the expression of INPP5E in control, NC, and si-SNHG12 groups. (e, f) RT-PCR assay was used to detect the mRNA expression of IFT88, Beclin1, MAP1LC3A, and MAP1LC3B, and WB assay was used to detect the protein expression of IFT88, Beclin1, LC3 I, and LC3 II in control, NC, mimic-1, and mimic-1 + OE-INPP5E groups. (g, h) RT-PCR assay was used to detect the mRNA expression of IFT88, Beclin1, MAP1LC3A, and MAP1LC3B, and WB assay was used to detect the protein expression of IFT88, Beclin1, LC3 I, and LC3 II in control, NC, mimic-2, and mimic-2 + OE-INPP5E groups. (i, j) RT-PCR assay was used to detect the mRNA expression of IFT88, Beclin1, MAP1LC3A, and MAP1LC3B, and WB assay was used to detect the protein expression of IFT88, Beclin1, LC3 I, and LC3 II in control, NC, si-SNHG12-1, and si-SNHG12-1 + OE-INPP5E groups. (k) Ace-tubulin was used to label primary cilia, and LC3 II was used to label autophagy via immunofluorescence assay. Notes: , , and compared with the control or between the indicated groups.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)