Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: MCC expression in podocytes in mouse and human. (A) MCC mRNA was detected in 18 out the 20 mouse podocytes subjected to single-cell RNA-seq analysis (GEO: GSE88814). (B) MCC expression in mouse podocytes, mesangial cells and glomerular endothelial cells from the bulk sequencing of mouse glomerular cells (GEO: GSE123179). (C) MCC expression in glomerular cell types according to the database, KIT ( http://humphreyslab.com/SingleCell/ ). (D) Immunohistochemical staining of MCC in a mouse glomerulus, showing a staining pattern characteristic of podocytes. (E) MCC expression in human glomerular cell types, podocytes (Pod), mesangial cells (MC) and endothelial cells (EC) according to the KIT database above. (F) The immunohistochemical staining of MCC from the Human Protein Atlas ( www.proteinatlas.org ) shows that MCC expression is relatively specific to podocytes. (G) Fluorescence co-immunostaining of MCC and SYNPO in mouse kidney, showing the localization of MCC in foot processes of podocytes.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Expressing, RNA Sequencing, Sequencing, Immunohistochemical staining, Staining, Fluorescence, Immunostaining

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: Subcellular localization of MCC protein in podocytes in culture. (A) Fluorescence immunostaining of MCC and DAPI in cultured podocytes treated with scramble and si-MCC, respectively, which shows MCC protein in the cytoplasm, nucleus and periphery of the cells. (B) Representative immunoblotting of MCC in the cytoplasmic and nuclear fractions of human glomerular cells. GAPDH served the cytoplasmic marker, while Lamin B1 the nuclear marker. MCC expression in tubules was also examined, showing cytoplasmic and nuclear distribution; however, the protein levels were both significantly lower than that in glomerular cell cytoplasm and nuclei, respectively.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Fluorescence, Immunostaining, Cell Culture, Western Blot, Marker, Expressing

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: Morphological change of podocytes treated with si-MCC. At 72 h post transfection of si-MCC, the podocytes exhibited an elongated shape compared with the scramble control cells.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Transfection, Control

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: MCC silencing resulted in injury in podocytes. (A) Phalloidin staining of the cells treated with scramble control and si-MCC for 72 h. (B) Quantification of the staining intensity using ImageJ showed a significant reduction of actin stress fibers in the cells treated with si-MCC compared with scramble control. (C) qPCR quantification of CD2AP mRNA in control cells and the cells treated with si-MCC at 36 h and 72 post transfection, respectively. (D) Immunoblotting of the CD2AP protein in the cell samples in C to confirm CD2AP downregulation at the protein level. Quantification of the blot is shown on the right. (E) qPCR analysis of WT1 in the cells treated with scramble control and si-MCC, respectively. (F) Immunoblotting of WT1 in the samples in (E) and the quantification shown on the right. The data represent the mean ± SD of three independent experiments. * p < 0.05, statistically significant; ** p < 0.01 and *** p < 0.001, statistically very significant.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Staining, Control, Transfection, Western Blot

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: Flow cytometry of Annexin V staining of podocytes treated with scramble control and si-MCC for 72h. (A) Histogram representation of the results of the flow cytometric assay. (B) Quantification of the percentages of the Annexin V positive cells in the samples. The results were obtained from three independent experiments ( n = 3) and calculated as mean ± SD. * p < 0.05, statistically significant.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Flow Cytometry, Staining, Control

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: GO_BP analyses of genes regulated in the podocytes deficient in MCC.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques:

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: GO_CC analyses of genes regulated in the podocytes deficient in MCC.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques:

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: Requirement of MCC in lamellipodium formation of podocytes. (A) Fluorescence immunostaining of MCC and F-actin (phalloidin) shows that MCC protein is abundant in lamellipodia (arrows), and its knockdown caused loss of lamellipodia (arrow head) in the podocytes. (B) Fluorescence immunostaining of lamellipodial marker, cortactin, further shows the loss of lamellipodia in the cells lacking MCC (arrow head).

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Fluorescence, Immunostaining, Knockdown, Marker

Journal: Frontiers in Medicine

Article Title: MCC Regulator of WNT Signaling Pathway (MCC) Is a Podocyte Essential Gene

doi: 10.3389/fmed.2021.777563

Figure Lengend Snippet: MCC downregulation in podocyte injury and podocytopathy. (A) qPCR analysis of cultured podocytes treated with or without PAN for 24 h, showing a significant downregulation of MCC by PAN. The results represent the mean ± SD of three independent experiments; *** p < 0.001, **** p < 0.0001, statistically very significant; (B) Nephroseq database search showed MCC downregulation in glomeruli of diabetic mice (eNOS-deficient C57BLKS db/db) ( n = 7) compared with the non-diabetic control mice ( n = 5). * p < 0.01; (C) Nephroseq database search revealed downregulation of MCC in glomeruli of FSGS patients. Normal control, n = 9; FSGS patients, n = 6; * p < 0.05, statistically significant.

Article Snippet: Consistently, from the Human Protein Atlas ( www.proteinatlas.org ), MCC was found mainly expressed in podocytes as shown by the immunohistochemical staining pattern characteristic of podocytes along the periphery of glomerular tuft ( ).

Techniques: Cell Culture, Control